Mannose derivatives for treating bacterial infections

ABSTRACT

The invention also provides pharmaceutically acceptable compositions containing the compounds and methods of using the compositions in the treatment of bacteria infections. Finally, the invention provides processes for making compounds of the invention.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a continuation application which claims priority toU.S. patent application Ser. No. 15/140,045, filed on Apr. 27, 2016,which is a divisional application which claims priority to U.S. patentapplication Ser. No. 14/132,662, filed on Dec. 18, 2013, which claimsthe benefit, under 35 U.S.C. § 119, of U.S. Provisional Application No.61/738,620, filed Dec. 18, 2012; U.S. Provisional Application No.61/788,241, filed Mar. 15, 2013; and U.S. Provisional Application No.61/874,501, filed Sep. 6, 2013, the entire contents of each of the aboveapplications being incorporated herein by reference.

DESCRIPTION OF THE FIGURES

FIG. 1: Differential scanning calorimetry (DSC) of Compound 162,crystalline form A

FIG. 2: X-ray powder diffractogram of Compound 162

FIG. 3: Differential scanning calorimetry (DSC) of Compound 202,crystalline form A

FIG. 4: X-ray powder diffractogram of Compound 202

FIG. 5: Thermal gravimetric analysis (TGA) trace of Compound 202

BACKGROUND OF THE INVENTION

Inflammatory bowel disease (IBD) is a complex chronic inflammatorydisorder, with the two more common forms being ulcerative colitis (UC)and Crohn's disease (CD). IBD is a multifactorial disease that resultsfrom a combination of predisposing genetic factors, environmentaltriggers, dysbiosis of the gastrointestinal microbiota and aninappropriate inflammatory response (Man et al., 2011, Nat RevGastroenterol Hepatol, March, 8(3):152-68).

Several studies on fecal and mucosa-associated bacterial communitieshave shown that the microbiota of patients with Crohn's disease (CD)differ from those of healthy controls, as well as those of patients withulcerative colitis (UC). Although the reported changes are not alwaysconsistent, numbers of Escherichia coli are generally increased, whereasFirmicutes are scarcer in CD patients (Peterson et al., 2008, Cell HostMicrobe, 3: 17-27; Frank et al., 2007, Proc. Natl. Acad. Sci.,104:13780-13785). Whether these changes are causative factors orconsequences of inflammation, it remains controversial. To date, severalpathogens have been proposed as causative agents. In particular,adherent-invasive E. coli (AIEC) has been reported to be more prevalentin CD patients than in controls in several countries (United Kingdom,France and the USA) (Darfeuille-Michaud et al., 2004, Gastroenterology,127:412-421; Martinez-Medina et al., 2009, Inflamm Bowel Dis.,15:872-882). AIEC strains have been isolated from ileal lesions in ˜35%of CD patients compared to ˜5% of healthy subjects. One of the featuresof AIEC is their ability to adhere and invade epithelial cells. It isknown from various models that the binding of adhesins expressed on thebacterial cell surface to defined glycosylated receptors on the hosttissue surface is considered to be an initial and critical step inpathogenesis, then opening a new avenue for therapy such as blocking theinteraction between type 1 pili and CEACAM6, a known host receptor forFimH (Barnich et al., 2007, J. Clin. Invest., 117:1566-1574; Carvalho etal., 2009, JEM, vol. 206, no. 10, 2179-2189). Therefore, inhibition ofadhesion, and consequently intracellular replication of AIEC inepithelial cells, may prevent establishment of a sub-mucosal infectionleading to mucosal inflammation and epithelial barrier disruption.

It has also been demonstrated recently that FimH antagonists arepotentially effective in treating urinary tract infections (J. Med.Chem. 2010, 53, 8627-8641).

SUMMARY OF THE INVENTION

The present invention provides compounds useful for the treatment orprevention of bacteria infections, such as urinary tract infection (UTI)and inflammatory bowel disease (IBD).

The compounds of the present invention are represented by the followingstructure of Formula (I), or a pharmaceutically acceptable salt thereof:

wherein V¹, Z, and V are as described herein.

The present invention also provides a composition comprising thecompound described herein, and a pharmaceutically acceptable carrier,adjuvant, or vehicle.

The present invention also provides a method of treating or preventingbacteria infection in a subject, comprising administering to the subjectan effective amount of the compound or the composition described herein.The present invention also provides processes for making compounds ofthe invention.

DETAILED DESCRIPTION OF THE INVENTION

The present invention relates to compounds useful for the treatment orprevention of bacteria infections, such as urinary tract infection (UTI)and inflammatory bowel diseases (IBD).

One aspect of the invention provides a compound of Formula I, or apharmaceutically acceptable salt thereof:

wherein

-   V¹ is halogen, NH₂, OH, or SH;-   V is H, halogen, —OR⁷, —NR⁵R⁶, —SR⁷, or C₁-C₆aliphatic;-   R⁵ is —H; X⁵; Q⁵; X⁵-Q⁵; —C(O)R⁹; —C(O)NHR⁹; or —C(O)OR⁹;-   R⁶ is —H; X⁶; Q⁶; X⁶-Q⁶; —C(O)R⁹; —C(O)NHR⁹; or —C(O)OR⁹;-   R⁷ is —H; X⁷; Q⁷; X⁷-Q⁷; —C(O)R⁹; or —C(O)NHR⁹;-   R⁹ is —H; X⁹; Q⁹; or X⁹-Q⁹;-   each X⁵, X⁶, X⁷, and X⁹ is independently C₁-C₆aliphatic optionally    substituted with 1-3 halo;-   each Q⁵, Q⁶, Q⁷, and Q⁹ is independently C₆-C₁₀ aryl, 5-10 membered    heteroaryl, C₃-C₈ cycloaliphatic, or 3-12 membered heterocyclyl;    wherein said Q⁵, Q⁶, Q⁷, and Q⁹ is independently and optionally    substituted with 1-6 occurrences of J;-   Z is

-   wherein Rings A, B, C, D, E, F, G, H, I, K, M, N, O, P, Q, R, S, T,    U, V, W, X, Y, and Z are each independently a 5-6 membered    saturated, fully unsaturated, partially unsaturated, or aromatic    monocyclic ring optionally having 1-4 heteroatoms selected from    oxygen, nitrogen, or sulfur; an 8-12 membered saturated, fully    unsaturated, partially unsaturated, or aromatic bicyclic ring    optionally having 1-6 heteroatoms selected from oxygen, nitrogen, or    sulfur; or a 10-14 membered saturated, fully unsaturated, partially    unsaturated, or aromatic tricyclic ring optionally having 1-6    heteroatoms selected from oxygen, nitrogen, or sulfur;-   L is —X    Y, wherein X is a C₁aliphatic or —C(O)— and Y is C₁-C₁₀aliphatic    wherein up to two methylene units of the C₁-C₁₀aliphatic are    optionally replaced with —C(O)—, NH, or NH(C₁₋₆aliphatic); L is    optionally substituted with 1-3 halo;-   L¹ is —X¹    Y¹— wherein X¹ is a C₁aliphatic or —C(O)— and Y¹ is C₁-C₁₀aliphatic    wherein up to two methylene units of the C₁-C₁₀aliphatic are    optionally replaced with —C(O)—, NH, or N(C₁-C₆aliphatic); L¹ is    optionally substituted with 1-3 halo;-   L² is —X²    Y²— wherein X² is a C₁aliphatic or —C(O)— and Y² is C₁-C₁₀aliphatic    wherein up to two methylene units of the C₁-C₁₀aliphatic are    optionally replaced with —C(O)—, NH, or N(C₁-C₆aliphatic); L² is    optionally substituted with 1-3 halo;-   each L³, L⁵, and L¹⁶ is independently C₁-C₁₂aliphatic wherein up to    three methylene units of the C₁-C₁₂aliphatic are optionally replaced    with —C(O)—, NH, or N(C₁-C₆aliphatic); each L³, L⁵, and L¹⁶ is    independently and optionally substituted with 1-3 halo;-   L⁴ is X⁴    Y⁴ wherein X⁴ is a C₁aliphatic or —C(O)— and Y⁴ is C₁-C₁₀aliphatic    wherein up to two methylene units of the C₁-C₁₀aliphatic are    optionally replaced with —C(O)—, NH, or N(C₁-C₆aliphatic); L⁴ is    optionally substituted with 1-3 halo;-   L⁶ is C₁-C₁₅aliphatic wherein up to six methylene units of the    C₁-C₁₅aliphatic are optionally replaced with O, NH,    N(C₁-C₆aliphatic), S, —C(O)—, S(O), or S(O)₂; L⁶ is optionally    substituted with 1-3 halo;-   each L⁷ and L⁹ is independently C₁-C₆aliphatic wherein up to two    methylene units of the C₁-C₆aliphatic are optionally replaced with    —C(O)—, NH, or N(C₁-C₄aliphatic); each L⁷ and L⁹ is independently    and optionally substituted with 1-3 halo;-   L⁸ is —X⁸    Y⁸— wherein X⁸ is a C₁aliphatic and Y⁸ is C₁-C₁₀aliphatic wherein up    to two methylene units of the C₁-C₁₀aliphatic are optionally    replaced with —C(O)—, NH, or N(C₁-C₆aliphatic); L⁸ is optionally    substituted with 1-3 halo;-   L¹⁰ is C₁-C₆aliphatic wherein up to two methylene units of the    C₁-C₆aliphatic are optionally replaced with O, NH,    N(C₁-C₆aliphatic), S, or —C(O)—; L¹⁰ is optionally substituted with    1-3 halo;-   L¹¹ is C₁-C₆aliphatic wherein up to two methylene units of the    C₁-C₆aliphatic are optionally replaced with O, NH,    N(C₁-C₆aliphatic), S, or —C(O)—; L¹¹ is optionally substituted with    1-3 halo;-   L¹² is C₁-C₆aliphatic wherein up to two methylene units of the    C₁-C₆aliphatic are optionally replaced with O, NH,    N(C₁-C₆aliphatic), S, or —C(O)—; L¹² is optionally substituted with    1-3 halo;-   L¹³ is C₁-C₆aliphatic wherein up to two methylene units of the    C₁-C₆aliphatic are optionally replaced with O, NH,    N(C₁-C₆aliphatic), S, or —C(O)—; L¹³ is optionally substituted with    1-3 halo;-   L¹⁴ is —X¹⁴—    Y¹⁴— wherein X¹⁴ is a C₁aliphatic and Y¹⁴ is C₁-C₁₀aliphatic wherein    up to two methylene units of the C₁-C₁₀aliphatic are optionally    replaced with —C(O)—, NH, or N(C₁-C₆aliphatic); L¹⁴ is optionally    substituted with 1-3 halo;-   L¹⁵ is C₁-C₆aliphatic;-   each J, J^(A), J^(B), J^(C), J^(D), J^(E), J^(F), J^(G), J^(H),    J^(I), J^(K), J^(M), J^(N), J^(O), J^(P), J^(Q), J^(R), J^(S),    J^(T), J^(U), J^(V), J^(X), J^(Y) and J^(Z) is independently    halogen, —CN, —NO₂, X^(J), Q^(J), or X^(J)-Q^(J); or two J, J^(A),    J^(B), J^(C), J^(D), J^(E), J^(F), J^(G), J^(H), J^(I), J^(K),    J^(M), J^(N), J^(O), J^(P), J^(Q), J^(R), J^(S), J^(T), J^(U),    J^(V), J^(W), J^(X), J^(Y) or J^(Z) groups bound to the same carbon    atom, together with the carbon atom to which they are bound,    optionally form —C═N—OH, —C(O)—, or Ring HH;-   Ring HH is a 3-8 membered saturated monocyclic ring having 0-2    heteroatoms selected from oxygen, nitrogen, or sulfur; optionally    substituted with 1-4 occurrences of J^(HH);-   J^(HH) is halo, CN, oxo, X^(J), Q^(J), or X^(J)-Q^(J);-   X^(J) is a C₁-C₁₀ aliphatic, wherein up to 4 methylene units of the    C₁-C₁₀aliphatic are optionally replaced with —O—, —NH,    N(C₁-C₆aliphatic), —S—, —C(O)—, —C(═NOH)—, —S(O)—, —S(O)₂—, P, or    P(O); X^(J) is optionally substituted with 0-6 occurrences of halo,    OH, or C₁₋₄alkyl; or optionally substituted with 0-1 occurrences of    CN;-   Q^(J) is a 3-7 membered monocyclic saturated, fully unsaturated,    partially unsaturated, or aromatic ring optionally having 1-4    heteroatoms selected from oxygen, nitrogen, or sulfur; or an 8-12    membered saturated, fully unsaturated, partially unsaturated, or    aromatic ring optionally having 1-6 heteroatoms selected from    oxygen, nitrogen, or sulfur; wherein each Q^(J) is optionally    substituted with 1-6 occurrences of halo, oxo, CN, or C₁₋₆alkyl,    wherein up to 2 methylene units of said C₁₋₆alkyl are optionally    replaced with —O—, —NH, N(C₁-C₆aliphatic), —S—, —C(O)—, —S(O)—, or    —S(O)₂—;-   provided that in some embodiments, Z is not CH₂CH₂,

It shall be understood that a and b denote the bonds and indicate thespecific connectivity and stereochemistry of the bond connecting thesugar ring to group Z. It shall further be understood that when X, X¹,X², X⁴, X⁸, and X¹⁴ are C(O), the bond between X and Y; X¹ and Y¹; X²and Y²; X⁴ and Y⁴; X⁸ and Y⁸ and X¹⁴ and Y¹⁴ respectively is a singlebond. When X, X¹, X², X⁴, X⁸, and X¹⁴ is a C₁ aliphatic, then the bondbetween X and Y; X¹ and Y¹; X² and Y²; X⁴ and Y⁴; X⁸ and Y⁸ and X¹⁴ andY¹⁴ respectively will change depending on the nature of X, X¹, X², X⁴,X⁸, and X¹⁴. For example, if X is “CH”, then the bond between X and Y isa double bond to result in —CH═Y.

Another aspect of the invention provides a compound of Formula I, or apharmaceutically acceptable salt thereof:

wherein

-   V¹ is halogen, NH₂, OH, or SH;-   V is H, halogen, —OR⁷, —NR⁵R⁶, —SR⁷, or C₁-C₆aliphatic;-   R⁵ is —H; X⁵; Q⁵; X⁵-Q⁵; —C(O)R⁹; —C(O)NHR⁹; or —C(O)OR⁹;-   R⁶ is —H; X⁶; Q⁶; X⁶-Q⁶; —C(O)R⁹; —C(O)NHR⁹; or —C(O)OR⁹;-   R⁷ is —H; X⁷; Q⁷; X⁷-Q⁷; —C(O)R⁹; or —C(O)NHR⁹;-   R⁹ is —H; X⁹; Q⁹; or X⁹-Q⁹;-   each X⁵, X⁶, X⁷, and X⁹ is independently C₁-C₆aliphatic optionally    substituted with 1-3 halo;-   each Q⁵, Q⁶, Q⁷, and Q⁹ is independently C₆-C₁₀ aryl, 5-10 membered    heteroaryl, C₃-C₈ cycloaliphatic, or 3-12 membered heterocyclyl;    wherein said Q⁵, Q⁶, Q⁷, and Q⁹ is independently and optionally    substituted with 1-6 occurrences of J;-   Z is

-   wherein Rings A, B, C, D, E, F, G, H, I, K, M, N, O, P, Q, R, S, T,    U, V, W, and X are each independently a 5-6 membered fully    unsaturated, partially unsaturated, or monocyclic aromatic ring    optionally having 1-4 heteroatoms selected from oxygen, nitrogen, or    sulfur; an 8-12 membered fully unsaturated, partially unsaturated,    or aromatic bicyclic ring optionally having 1-6 heteroatoms selected    from oxygen, nitrogen, or sulfur; or a 10-14 membered fully    unsaturated, partially unsaturated, or aromatic tricyclic ring    optionally having 1-6 heteroatoms selected from oxygen, nitrogen, or    sulfur;-   L is —X    Y, wherein X is a C₁aliphatic or —C(O)— and Y is C₁-C₁₀aliphatic    wherein up to two methylene units of the C₁-C₁₀aliphatic are    optionally replaced with —C(O)—, NH, or NH(C₁₋₆aliphatic); L is    optionally substituted with 1-3 halo;-   L¹ is —X¹    Y¹— wherein X¹ is a C₁aliphatic or —C(O)— and Y¹ is C₁-C₁₀aliphatic    wherein up to two methylene units of the C₁-C₁₀aliphatic are    optionally replaced with —C(O)—, NH, or N(C₁-C₆aliphatic); L¹ is    optionally substituted with 1-3 halo;-   L² is —X²    Y² wherein X² is a C₁aliphatic or —C(O)— and Y² is C₁-C₁₀aliphatic    wherein up to two methylene units of the C₁-C₁₀aliphatic are    optionally replaced with —C(O)—, NH, or N(C₁-C₆aliphatic); L² is    optionally substituted with 1-3 halo;-   each L³ and L⁵ is independently C₁-C₁₂aliphatic wherein up to three    methylene units of the C₁-C₁₂aliphatic are optionally replaced with    —C(O)—, NH, or N(C₁-C₆aliphatic); each L³ and L⁵ is independently    and optionally substituted with 1-3 halo;-   L⁴ is —X⁴    Y⁴— wherein X⁴ is a C₁aliphatic or —C(O)— and Y⁴ is C₁-C₁₀aliphatic    wherein up to two methylene units of the C₁-C₁₀aliphatic are    optionally replaced with —C(O)—, NH, or N(C₁-C₆aliphatic); L⁴ is    optionally substituted with 1-3 halo;-   L⁶ is C₁-C₁₅aliphatic wherein up to six methylene units of the    C₁-C₁₅aliphatic are optionally replaced with O, NH,    N(C₁-C₆aliphatic), S, —C(O)—, S(O), or S(O)₂; L⁶ is optionally    substituted with 1-3 halo;-   each L⁷ and L⁹ is independently C₁-C₆aliphatic wherein up to two    methylene units of the C₁-C₆aliphatic are optionally replaced with    —C(O)—, NH, or N(C₁-C₄aliphatic); each L⁷ and L⁹ is independently    and optionally substituted with 1-3 halo;-   L⁸ is —X⁸    Y⁸ wherein X⁸ is a C₁aliphatic and Y⁸ is C₁-C₁₀aliphatic wherein up    to two methylene units of the C₁-C₁₀aliphatic are optionally    replaced with —C(O)—, NH, or N(C₁-C₆aliphatic); L⁸ is optionally    substituted with 1-3 halo;-   L¹⁰ is C₁-C₆aliphatic wherein up to two methylene units of the    C₁-C₆aliphatic are optionally replaced with O, NH,    N(C₁-C₆aliphatic), S, or —C(O)—; L¹⁰ is optionally substituted with    1-3 halo;-   L¹¹ is C₁-C₆aliphatic wherein up to two methylene units of the    C₁-C₆aliphatic are optionally replaced with O, NH,    N(C₁-C₆aliphatic), S, or —C(O)—; L¹¹ is optionally substituted with    1-3 halo;-   L¹² is C₁-C₆aliphatic wherein up to two methylene units of the    C₁-C₆aliphatic are optionally replaced with O, NH,    N(C₁-C₆aliphatic), S, or —C(O)—; L¹¹ is optionally substituted with    1-3 halo;-   L¹³ is C₁-C₆aliphatic wherein up to two methylene units of the    C₁-C₆aliphatic are optionally replaced with O, NH,    N(C₁-C₆aliphatic), S, or —C(O)—; L¹¹ is optionally substituted with    1-3 halo;-   each J, J^(A), J^(B), J^(C), J^(D), J^(E), J^(F), J^(G), J^(H),    J^(I), J^(K), J^(M), J^(N), J^(O), J^(P), J^(Q), J^(R), J^(S),    J^(T), J^(U), J^(V), J^(W) and J^(X) is independently halogen, —CN,    —NO₂, X^(J), Q^(J), or X^(J)-Q^(J);-   X^(J) is a C₁-C₁₀ aliphatic, wherein up to 4 methylene units of the    C₁-C₁₀ aliphatic are optionally replaced with —O—, —NH,    N(C₁-C₆aliphatic), —S—, —C(O)—, —C(═NOH)—, —S(O)—, —S(O)₂—, P, or    P(O); X^(J) is optionally substituted with 0-6 occurrences of halo;-   Q^(J) is a 3-7 membered monocyclic fully unsaturated, partially    unsaturated, or aromatic ring optionally having 1-4 heteroatoms    selected from oxygen, nitrogen, or sulfur; or an 8-12 membered fully    unsaturated, partially unsaturated, or aromatic ring optionally    having 1-6 heteroatoms selected from oxygen, nitrogen, or sulfur;    wherein each Q^(J) is optionally substituted with 1-6 occurrences of    halogen, CN, NO₂, C₁-C₆aliphatic, OH, NH₂, NH(C₁-C₆aliphatic),    NH(C₁-C₆aliphatic)₂, phenyl, 5-6 membered heteroaryl, C₃-C₆    cycloaliphatic, or 3-8 membered heterocyclyl, wherein said phenyl,    5-6 membered heteroaryl, C₃-C₆ cycloaliphatic, or 3-8 membered    heterocyclyl is optionally substituted with halo, CN, NO₂, or    C₁-C₆aliphatic wherein up to 3 methylene units of the C₁-C₆aliphatic    are optionally replaced with O, NH, S, or CO;-   provided that Z is not CH₂CH₂,

In some embodiments, the C₆-C₁₀ aryl of Q⁵, Q⁶, Q⁷, and Q⁹ is phenyl ornapthyl; the 5-10 membered heteroaryl is a 5-6 membered monocyclicheteroaryl having 1-4 heteroatoms selected from oxygen, nitrogen, orsulfur; or an 8-10 bicyclic heteroaryl having 1-4 heteroatoms selectedfrom oxygen, nitrogen, or sulfur; the C₃-C₈ cycloaliphatic is amonocyclic C₃-C₈ cycloalkyl or cycloalkenyl ring; and the 3-12 memberedheterocyclyl is a 3-8 membered monocyclic heterocyclyl having 1-3heteroatoms selected from oxygen, nitrogen, or sulfur; or an 8-12membered bicyclic heterocyclyl having 1-6 heteroatoms selected fromoxygen, nitrogen, or sulfur.

Some embodiments comprise one or more of the following:

-   -   a) Z is selected from formula i, ii, iii, iv, v, vi, vii, viii,        ix, x, xi, or xii;    -   b) Rings A, B, C, D, E, F, G, H, I, K, M, N, O, P, Q, R, S, T,        U, V, W, and X are each independently a 5-6 membered fully        unsaturated, partially unsaturated, or aromatic monocyclic ring        optionally having 1-4 heteroatoms selected from oxygen,        nitrogen, or sulfur; or an 8-12 membered fully unsaturated,        partially unsaturated, or aromatic bicyclic ring optionally        having 1-6 heteroatoms selected from oxygen, nitrogen, or        sulfur; and    -   c) X^(J) is a C₁-C₁₀ aliphatic, wherein up to 4 methylene units        of the C₁-C₁₀ aliphatic are optionally replaced with —O—, —NH,        N(C₁-C₆aliphatic), —S—, —C(O)—, —C(═NOH)—, —S(O)—, —S(O)₂—, P,        or P(O); X^(J) is optionally substituted with 0-6 occurrences of        halo.

In some embodiments, V¹ is OH. In other embodiments, V is OH.

In some embodiments, Ring A, B, C, D, E, F, G, H, I, K, M, N, O, P, Q,R, S, T, U, V, W, X, Y, and Z are aromatic.

In another embodiment, Z is selected from formula ii, iii, v, vii, orxiii. In some embodiments, Z is selected from formula ii, iii, or v.

In some embodiments, Ring A, B, C, D, E, F, G, H, I, K, M, N, O, P, Q,R, S, T, U, V, W, and X are aromatic. In other embodiments, Ring A, B,C, D, E, F, G, H, I, K, M, N, O, P, Q, R, S, T, U, V, W, X, Y, and Z areeach independently phenyl, napthyl, or a 5-6 membered heteroaryl having1-4 heteroatoms selected from oxygen, nitrogen, or sulfur. In yet otherembodiments, Ring A, B, C, D, E, F, G, H, I, K, M, N, O, P, Q, R, S, T,U, V, W, and X are each independently phenyl, napthyl, or a 5-6 memberedheteroaryl having 1-4 heteroatoms selected from oxygen, nitrogen, orsulfur. In some embodiments, each Ring A, B, D, M, O, T, and U is bondedto the mannose ring to which it is attached via a carbon atom.

In some embodiments, each J, J^(A), J^(B), J^(C), J^(D), J^(E), J^(F),J^(G), J^(H), J^(I), J^(K), J^(M), J^(N), J^(O), J^(P), J^(Q), J^(R),J^(S), J^(T), J^(U), J^(V), J^(W), J^(X), J^(Y) and J^(Z) isindependently —NO₂, —CN, halogen, or C₁-C₁₀aliphatic wherein up to threemethylene units of the C₁-C₁₀aliphatic is optionally replaced with O,NH, N(C₁₋₄alkyl), S, C(O), S(O), or S(O)₂ and optionally substitutedwith 1-3 halo or 1 CN. In other embodiments, each J, J^(A), J^(B),J^(C), J^(D), J^(E), J^(F), J^(G), J^(H), J^(I), J^(K), J^(M), J^(N),J^(O), J^(P), J^(Q), J^(R), J^(S), J^(T), J^(U), J^(V), J^(W), and J^(X)is independently —NO₂, —CN, halogen, or C₁-C₁₀aliphatic wherein up tothree methylene units of the C₁-C₁₀aliphatic is optionally replaced withO, NH, N(C₁₋₄alkyl), S, C(O), S(O), or S(O)₂ and optionally substitutedwith 1-3 halo.

In one aspect of the invention, Z is

In some embodiments, Ring A is triazolyl, thienyl, or phenyl and J^(A)is CF₃ or —O(C₁₋₆alkyl). In other embodiments, Ring A is phenyl andJ^(A) is CF₃ or OCH(CH₃)₂.

According to another aspect Z is

In some embodiments, Ring B and Ring C are independently triazolyl orphenyl. In other embodiments, Ring B and Ring C are phenyl. In someembodiments, J^(B) and J^(C) are each independently halo, C₁₋₆alkyl, orO(C₁₋₆alkyl).

According to another aspect Z is

In some embodiments, Ring D, Ring E, and Ring F are each independentlytriazolyl or phenyl. In other embodiments, Ring D and Ring E are phenyland Ring E is an optionally substituted group selected fromC₃₋₆cycloalkyl, phenyl, pyridinyl, or pyrazinyl. In some embodiments,Ring E is pyridinyl. In other embodiments, J^(D), J^(E), and J^(F) areeach independently C₁-C₆aliphatic wherein up to two methylene units ofthe C₁-C₆aliphatic is optionally replaced with O, NH, N(C₁₋₄alkyl), orC(O). In some embodiments, these substituents are C₁-C₆alkyl,O(C₁-C₆alkyl), halo, or CH₂C(O)OCH₃. In other embodiments, thesesubstituents are CH₃, OCH₃, fluoro, or CH₂C(O)OCH₃.

According to another aspect Z is

In some embodiments, L¹ is —C₁-C₆aliphatic and Ring G phenyl. In otherembodiments, Ring G is phenyl or indolyl; and J^(G) is C₁₋₆alkyl, halo,or —O(C₁₋₆alkyl). In another embodiment, L¹ is O. In other embodiments,L¹ is —C≡C—.

According to another aspect Z is

In some embodiments,

-   L² is C₁₋₆aliphatic or —(C₁₋₄aliphatic)-C(O)NH—;-   L³ is C₁₋₆aliphatic or —C(O)NH—(C₁₋₄aliphatic)-;-   Ring H is phenyl or naphthyl; and-   J^(H) is halo, CN, NO₂, C₁₋₆aliphatic, —OC₁₋₆aliphatic, or    C(O)O(C₁₋₆aliphatic).

In other embodiments,

-   L² is C₁₋₆aliphatic or —(C₁₋₄aliphatic)-C(O)NH—;-   L³ is C₁₋₆aliphatic or —NHC(O)—(C₁₋₄aliphatic)-;-   Ring H is phenyl or naphthyl; and-   J^(H) is halo, CN, NO₂, C₁₋₆aliphatic, —OC₁₋₆aliphatic, or    C(O)O(C₁₋₆aliphatic), wherein said J^(H) is optionally substituted    with 1-3 occurrences of halo.

In yet other embodiments, J^(H) is halo, CN, NO₂, phenyl, orC₁₋₁₀aliphatic wherein up to 3 methylene units are optionally replacedwith O, NH, N(C₁₋₄alkyl), S, C(O), SO, or SO₂; wherein said J^(H) isoptionally substituted with 1-3 occurrences of CN, halo or phenyl.

According to another aspect Z is

In some embodiments,

-   L⁴ and L⁵ are each independently C₁₋₆aliphatic;-   Ring I and Ring K are each independently phenyl;-   J^(I) and J^(K) are each independently halo, CN, NO₂, C₁₋₆aliphatic,    —OC₁₋₆aliphatic, or C(O)O(C₁₋₆aliphatic).

In another embodiment, L⁴ is —CH₂CH═CH— and L⁵ is —C≡C—.

According to another aspect Z is

In some embodiments, L⁶ is C₁-C₁₅aliphatic wherein up to four methyleneunits of the C₁-C₁₅aliphatic are optionally replaced with —O— or C(O)NH.In other embodiments, L⁶ is —C≡C—C≡C—, —C(O)NH—, —NHC(O)NH—,—(C₁₋₆alkyl)-, —C(O)NH—(C₁₋₈alkyl)-NHC(O)—.—C(O)NH—(CH₂CH₂)—O—(CH₂CH₂)—O—(CH₂CH₂)—NHC(O)—, or —CH₂N(CH₂C≡CH)CH₂—.In yet other embodiment, M is phenyl; N is phenyl; and each J^(M) andJ^(N) is each independently H or C₁₋₆alkyl.

Another aspect provides compounds wherein Z is

In some embodiments, Ring O, Ring P, and Ring Q are each independentlyphenyl, triazolyl, or thienyl. In some embodiments, L⁷ is —C(O)NH— orC₁₋₄alkyl. In other embodiments, Ring O, Ring P, and Ring Q are phenyl.

Another aspect provides compounds wherein Z is

In some embodiments, Ring R and Ring S are phenyl and L⁸ is C₁-C₆alkyl.In other embodiments, Ring R and Ring S are thienyl. In someembodiments, J^(R) is C₁₋₆alkyl.

According to another aspect Z is

In some embodiments, Ring T is phenyl or naphthyl and L¹⁰ isC₁-C₆aliphatic wherein up to one methylene unit of the C₁-C₆aliphatic isoptionally replaced with —O—. In some embodiments, L¹⁰ is —C≡C— or—CH₂CH₂—.

According to another aspect Z is

In some embodiments, L¹¹, L¹², and L¹³ are each independently —C(O)NH—or C₁₋₄alkyl and Ring U, V, W, and X are each independently phenyl.

According to another aspect Z is

In some embodiments, L is C₃-C₆ aliphatic. In some embodiments, L is anoptionally substituted C₁₋₆aliphatic. In other embodiments, L is —C≡C—C≡C—.

According to another aspect Z is

In some embodiments, Ring Y and Ring Z are phenyl. In some embodiments,L¹⁴ and L¹⁶ are C≡C—. In some embodiments, L¹⁵ is C₁₋₄aliphatic. In someembodiments, L¹⁵ is —C(CH₃)₂—. In certain embodiments, Ring Y and Ring Zare phenyl; L¹⁴ and L¹⁶ are C≡C—; and L¹⁵ is-C(CH₃)₂—.

According to another aspect, L¹, L², L³, L⁶, and L¹⁰ are eachindependently C₁-C₄ alkenyl or C₁-C₄ alkynyl.

Another aspect provides a compound having formula IA:

Another aspect provides a compound having formula IB:

In some embodiments, L² and L³ are bonded to the mannose ring via acarbon atom. In some embodiments, L² and L³ are each independently C₁-C₆alkenyl or C₁-C₆ alkynyl. In some embodiments at least one of L² and L³is —C≡C—. In other embodiments, L² and L³ are both —C≡C—.

Another aspect provides a compound having formula IC:

In some embodiments, L⁶ is bonded to the meta or para position of thephenyl ring(s) to which it is attached. In other embodiments, L⁶ isbonded to the para position of the phenyl ring(s) to which it isattached. In yet other embodiments, L⁶ is bonded to the meta position ofthe phenyl ring(s) as shown in Formula IC-a:

In some embodiments L⁶ is —C≡C—C≡C—. In other embodiments, L⁶ isC₁₋₆aliphatic, —O—(C₁₋₄alkyl)-O—, —C(O)NH—, —NHC(O)NH—,—C(O)NH—(C₁₋₁₀alkyl)-NHC(O)—, —(CH₂CH₂OCH₂CH₂OCH₂CH₂)—. In yet otherembodiments, L⁶ is —CH₂— or —C(CH₃)₂—.

Another aspect provides a compound having formula ID:

In some embodiments, Ring H is an optionally substituted 5-6 memberedmonocyclic aromatic ring optionally having 1-4 heteroatoms selected fromoxygen, nitrogen, or sulfur; or an 8-12 membered bicyclic aromatic ringoptionally having 1-6 heteroatoms selected from oxygen, nitrogen, orsulfur; or a 10-14 tricyclic aromatic ring optionally having 1-6heteroatoms selected from oxygen, nitrogen, or sulfur. For the sake ofclarity, a bicyclic or tricyclic ring is considered an aromatic ring ifit contains at least one aromatic ring.

In other embodiments, Ring H is optionally substituted phenyl, naphthyl,thienyl, isoxazolyl, pyridinyl, pyrazinyl, thienylthiophenyl,quinolinyl, quinazolinyl, benzothiadiazolyl, or fluorenyl. In otherembodiments, Ring H is optionally substituted phenyl or naphthyl.

According to another embodiment, Ring H, together with J^(H) and J^(HH),is selected from the following:

In some embodiments, J^(H) is halogen, oxo, CN, X^(J), Q^(J), orX^(J)-Q^(J); wherein

-   X^(J) is C₁-C₁₀ aliphatic, wherein up to 4 methylene units of the    C₁-C₁₀ aliphatic are optionally replaced with —O—, —NH,    N(C₁-C₆aliphatic), —S—, —C(O)—, —S(O)—, —S(O)₂—;-   Q^(J) is phenyl; and-   J^(H) is optionally substituted with 0-3 occurrences of halo or 0-1    occurrences of CN.

In some embodiments, Ring H is optionally substituted phenyl ornaphthyl.

In other embodiments, J^(H) is halogen, CN, —C(CH₃)₂CN, C₃₋₆cycloalkyl,phenyl, —O—CH₂phenyl, or C₁₋₆alkyl wherein up to one methylene unit isoptionally replaced with —O—, —S—, —NH—, —N(C₁₋₆alkyl)-, or —C(O)—. Inyet other embodiments, Ring H is phenyl and J^(H) is halo, CN,—C(CH₃)₂CN, C₃₋₆cycloalkyl, phenyl, CH₂phenyl, —O—CH₂phenyl, orC₁₋₆alkyl wherein up to one methylene unit is optionally replaced with—O—, —S—, —NH—, —N(C₁₋₆alkyl)-, or —C(═O)—. In some embodiments, J^(H)is substituted with 0-3 halo or 0-1 CN.

According to another embodiment, Ring H is

-   wherein G is O, S, S(O), S(O)₂, CF₂, C(J^(H1))(J^(H2)),    —C(J^(H3))₂-C(J^(H4))₂-, or N(J^(H5));-   J^(H1) is H, OH, or C₁₋₆alkyl wherein up to 2 methylene units are    optionally replaced with —O—, —NH—, —NH(C₁-C₆aliphatic)-, —S—,    —C(O)—, —S(O)—, or —S(O)₂—; J^(H1) is optionally and independently    substituted with 1-3 occurrences of OH;-   J^(H2) is X^(JH), Q^(JH), or X^(JH)-Q^(JH); J^(H2) is optionally    substituted with 1-3 occurrences of OH;-   X^(JH) is C₁₋₆alkyl wherein up to 3 methylene units of C₁₋₆alkyl is    optionally replaced with —O—, —NH, N(C₁-C₆aliphatic), —S—, —C(O)—,    —S(O)—, or —S(O)₂—;-   Q^(JH) is C₃₋₆cycloalkyl, phenyl, or a 5-7 membered monocyclic    heterocyclyl having 1-3 heteroatoms selected from oxygen, nitrogen,    or sulfur;-   or J^(H1) and J^(H2), together with the carbon atom to which they    are attached, form C═N—OH, C═O, or Ring HH;-   Ring HH is a 5-7 membered saturated monocyclic ring having 0-2    heteroatoms selected from oxygen, nitrogen, or sulfur; wherein said    ring is optionally substituted with 1-4 occurrences of J^(HH);-   J^(HH) is halo, CN, X^(J), Q^(J), or X^(J)-Q^(J);-   J^(H5) is X^(J), Q^(J), or X^(J)-Q^(J);-   X^(J) is a C₁-C₁₀ aliphatic, wherein up to 4 methylene units of the    C₁-C₁₀ aliphatic are optionally replaced with —O—, —NH,    N(C₁-C₆aliphatic), —S—, —C(O)—, —S(O)—, or —S(O)₂—; X^(J) is    optionally substituted with 0-6 occurrences of halo or 0-1    occurrences of CN;-   Q^(J) is a 3-6 membered saturated, partially unsaturated, or    aromatic monocyclic ring optionally having 1-4 heteroatoms selected    from oxygen, nitrogen, or sulfur; or an 8-12 membered saturated,    partially unsaturated, or aromatic bicyclic ring optionally having    1-6 heteroatoms selected from oxygen, nitrogen, or sulfur; wherein    each Q^(J) is optionally substituted with 1-6 occurrences of    halogen, oxo, CN, NO₂ or a C₁-C₆aliphatic wherein up to 3 methylene    units of the C₁-C₆aliphatic are optionally replaced with O, NR, S,    or CO;-   each J^(H) J^(H3) and J^(H4) is independently H, halo, CN, or C₁-C₁₀    aliphatic, wherein up to 3 methylene units of the C₁-C₁₀ aliphatic    are optionally replaced with —O—, —NH, N(C₁-C₆aliphatic), S, —C(O)—,    —S(O)—, or SO₂—; each J^(H), J^(H3) and J^(H4) is independently and    optionally substituted with 0-2 occurrences of halo, OH, or    C₁₋₄alkyl or with 1 occurrence of CN; and-   R is H or C₁₋₄alkyl.    In some embodiments,-   G is C(J^(H1))(J^(H2));-   J^(H1) OH, F, or —CH₂CH₂OH;-   J^(H2) is OH, CH₃, cyclopropyl, F, CH₂CH₃, —CH₂CH₂OH,    —CH₂CH(OH)CH₂OH, or phenyl optionally substituted with OCH₃;-   or J^(H1) and J^(H2), together with the carbon atom to which they    are attached, form ═N—OH or a 6-membered saturated monocyclic ring    having 0-2 heteroatoms selected from oxygen, nitrogen, or sulfur;    wherein said ring is optionally substituted with C₁₋₆alkyl, OH, NH₂,    —C(O)OCH₃, —C(O)OC(CH₃)₃, —C(O)C(CH₃)₂OH, or —S(O)₂CH₃.

In some embodiments, Ring HH is selected from cyclopentyl, cyclohexyl,piperidinyl, piperazinyl, 1,3-dithianyl, or tetrahydropyranyl. In someembodiments, X^(JH) is C₁₋₆alkyl and Q^(JH) is C₃₋₆cycloaliphatic,oxetanyl, tetrahydropyrrolidinyl, piperidinyl, piperazinyl, ormorpholinyl.

According to another embodiment, Ring H, together with Ring HH, isselected from one of the following formulae:

According to another embodiment, Ring H is H2:

In some embodiments,

-   J^(H5) is X^(J), Q^(J), or X^(J)-Q^(J); wherein-   X^(J) is a C₁-C₁₀ aliphatic, wherein up to 4 methylene units of the    C₁-C₁₀ aliphatic are optionally replaced with —O—, —NH,    N(C₁-C₆aliphatic), or —S—; X^(J) is optionally substituted with 1-2    occurrences of halo; and-   Q^(J) is a monocyclic 3-6 membered saturated monocyclic ring having    1-2 heteroatoms selected from oxygen, nitrogen, or sulfur; Q^(J) is    optionally substituted with 1-4 occurrences of halo, CN, NO₂, oxo,    or C₁-C₆aliphatic wherein up to 3 methylene units of the    C₁-C₆aliphatic are optionally replaced with O, NR, S, or CO.

In some embodiments, J^(H5) is H, phenyl, CH₂CH₂OH, CH₂CH₂OD₃,CH₂C(O)OH, CH₂C(O)OCH₂CH₃, CH₂C(O)N(CH₃)₂, CH₂CH(OH)CH₂OH,CH₂CH(OH)CH₂N(CH₃)₂,

According to another embodiment, Ring H is H1;

In some embodiments,

-   J^(H1) is H, OH, —(C₁₋₄alkyl)OH, or —(C₁₋₄alkyl)OC(O)(C₁₋₄alkyl);-   J^(H2) is X^(JH), Q^(JH), or X^(JH)-Q^(JH); wherein-   X^(JH) is C₁₋₆alkyl wherein up to 3 methylene units of C₁₋₆alkyl is    optionally replaced with —O—, —NH, N(C₁-C₆aliphatic), or —C(O)—;-   Q^(JH) is C₃₋₆cycloalkyl, phenyl optionally substituted with    —O(C₁₋₄alkyl), or piperazinyl optionally substituted with C₁₋₄alkyl.    In other embodiments,-   J^(H1) is H, OH, CH₂OH, CH₂CH₂OH, or CH₂OC(O)CH₃;-   J^(H2) is H, CH₂CH₃, CH₂CH₂CH₃, CH(CH₃)₂, CH₂CH₂CH(CH₃)₂, CH₂OH,    CH₂CH₂OH, CH₂CH₂C(O)OH, CH₂CH(OH)CH₂OH, CH₂OC(O)CH₃,    N(CH₃)CH₂CH₂N(CH₃)₂, phenyl, 3-methoxyphenyl, 4-methylpiperazinyl,    or CH₂-cyclohexyl; and-   J^(H) is absent (i.e. J^(H) is H or J^(H) is absent).

In yet other embodiments, X^(JH) is C₁₋₆alkyl and Q^(JH) isC₃₋₆cycloaliphatic, oxetanyl, tetrahydropyrrolidinyl, piperidinyl,piperazinyl, or morpholinyl.

Another embodiment provides a compound having formula ID-a:

In some embodiments,

-   Ring HH is a 3-8 membered saturated monocyclic ring having 0-2    heteroatoms selected from oxygen, nitrogen, or sulfur;-   J^(HH) is X^(J), Q^(J), or X^(J)-Q^(J);-   X^(J) is a C₁-C₁₀ aliphatic, wherein up to 4 methylene units of the    C₁-C₁₀ aliphatic are optionally replaced with —O—, —NH,    N(C₁-C₆aliphatic), —S—, —C(O)—, —S(O)—, or —S(O)₂—; X^(J) is    optionally substituted with 0-6 occurrences of halo or 0-1    occurrences of CN;-   Q^(J) is a 3-7 membered monocyclic saturated, partially unsaturated,    or aromatic ring optionally having 1-4 heteroatoms selected from    oxygen, nitrogen, or sulfur; wherein each Q^(J) is optionally    substituted with 1-6 occurrences of halogen, CN, NO₂, or    C₁-C₆aliphatic wherein up to three methylene units are optionally    replaced with O, NH, NH(C₁-C₆aliphatic), S, C(O), S(O), or S(O)₂;    and-   J^(H) is halogen, CN, NO₂, or C₁-C₆aliphatic wherein up to three    methylene units are optionally replaced with O, NH,    NH(C₁-C₆aliphatic), S, C(O), S(O), or S(O)₂.

In some embodiments, Ring HH is cyclopentyl, cyclohexyl,tetrahydropyranyl, 1,3 dithianyl, piperazinyl, piperidinyl, or oxepanyl.In other embodiments, Ring HH is piperidinyl or tetrahydropyranyl.

Another embodiment provides a compound having formula ID-b:

In some embodiments,

-   J^(HH) is X^(J), Q^(J), or X^(J)-Q^(J);-   X^(J) is a C₁-C₄ aliphatic, wherein up to two methylene units of the    C₁-C₄ aliphatic are optionally replaced with —O—, —NH,    N(C₁-C₆aliphatic), —S—, —C(O)—, —S(O)—, or —S(O)₂—;-   Q^(J) is a 3-6 membered monocyclic saturated, partially unsaturated,    or aromatic ring optionally having 1-4 heteroatoms selected from    oxygen, nitrogen, or sulfur; wherein each Q^(J) is optionally    substituted with 1-3 occurrences of halogen, CN, or C₁-C₆aliphatic    wherein up to two methylene units of said C₁-C₆aliphatic are    optionally replaced with O, NH, NH(C₁-C₆aliphatic), S, C(O), S(O),    or S(O)₂; and-   J^(H) is halogen or C₁₋₄alkyl.

In some embodiments, J^(HH) is H, C(O)(C₁₋₆alkyl), C(O)O(C₁₋₆alkyl),S(O)₂(C₁₋₆alkyl), C(O)(C₃₋₆cycloalkyl), C(O)(3-6 membered heterocyclyl),C(O)(5-6 membered heteroaryl), C(O)—(C₁₋₄alkyl)-(5-6 memberedheteroaryl), C(O)—(C₁₋₄alkyl)-(heterocyclyl); wherein said heteroaryl orheterocyclyl has 1-3 heteroatoms selected from oxygen, nitrogen, orsulfur; J^(HH) is optionally substituted with OH, O(C₁₋₆alkyl), oxo,C₁₋₆alkyl, CN, or halo.

In other embodiments, J^(HH) is H, C(O)CH₃, C(O)OC(CH₃)₃, C(O)OCH(CH₃)₂,C(O)OCH₂CH₃, C(O)OC(OH)(CH₃)₂, S(O)₂CH₃, C(O)CH(CH₃)₂, C(O)C(CH₃)₃,C(O)CH(CH₃)OCH₃,

Another embodiment provides a compound represented by a structuralformula selected from the group consisting of:

or a pharmaceutically acceptable salt thereof.

Another embodiment provides a compound represented by a structuralformula selected from the group consisting of:

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or a pharmaceutically acceptable salt thereof.

Another embodiment provides a compound represented by a structuralformula selected from the group consisting of:

or a pharmaceutically acceptable salt thereof.In some embodiments, the compound is compound 162:

In other embodiments, the compound is compound 202:

In other embodiments, the compound is compound 53:

Another embodiment provides processes for preparing the compounds ofthis invention.

One embodiment provides a process for preparing a compound of formula

wherein Ring H and J^(H) are as defined herein; comprising reacting acompound of formula XX:

wherein Hal is a halogen, such as bromo or iodo; and J^(H) is as definedherein; with Intermediate M:

under Sonogashira coupling conditions to form the compound of formulaXXII.

Another embodiment provides a process for preparing a compound offormula

wherein Ring H and J^(H) are as defined herein; comprising

-   -   a) reacting a compound of formula XX:

wherein Hal is a halogen, such as bromo or iodo; and J^(H) is as definedherein; with Intermediate P:

under Sonogashira coupling conditions;

-   -   b) deprotecting Intermediate P under acidic conditions (such as        TFA, THF, H₂O) or with TBAF to afford a compound of formula        XXII.        Another embodiment provides a process of preparing a compound of        formula IIXX comprising    -   a) reacting a compound of formula XXIII:

under Lewis acid catalyzed double Ferrier type alkynylation conditionsto afford a compound of formula XXIV:

-   -   b) performing a stereospecific bis-dihydroxylation of compound        XXIV followed by a saponification to form a compound of formula        XXII.

In some embodiments, Ring H is unsubstituted phenyl and Hal is iodo.

Another embodiment provides a process for making compound 202:

comprising reacting Intermediate M:

with Intermediate AG8:

under Sonogashira coupling conditions to form Compound 202.

Another embodiment provides a process for preparing Intermediate AG8,comprising the steps of:

-   -   a) reacting

in the presence of a suitable base, such as NaH, and a suitable solvent,such as THF; to form Intermediate AG5:

-   -   b) Reacting Intermediate AG5 under acidic conditions, such as        HCl in dioxane, to form Intermediate AG4:

-   -   c) Reacting Intermediate AG4 with acetyl chloride, a suitable        base (such as triethylamine), and a suitable solvent (such as        DMF), to form Intermediate AG8.

Another embodiment provides a process for preparing Compound 162

comprising reacting Intermediate M:

under Sonogashira coupling conditions to form Compound 162.

Another embodiment provides a process for purifying crude compound 162formed by the Sonogashira coupling conditions described above comprisingone or more of the following steps: acetylating Compound 162 undersuitable acetylating reaction conditions to form a compound of formula162AC:

Comprising the steps of

-   -   a) purifying compound 162AC via known purification methods; and    -   b) reacting compound 162AC with suitable deprotection conditions        to afford Compound 162. Examples of deprotection methods        include, for example, column chromatography.

The present invention also provides a composition comprising thecompound described herein, and a pharmaceutically acceptable carrier,adjuvant, or vehicle.

The present invention also provides a method of treating or preventingbacteria infection in a subject, comprising administering to the subjectan effective amount of the compound or the composition described herein.

In an embodiment of the method, the bacteria infection is urinary tractinfection or inflammatory bowel disease.

Another embodiment provides a method of treating or preventing abacteria infection in a subject, comprising administering to the subjectan effective amount of a compound described herein or a pharmaceuticallyacceptable salt thereof, or a composition comprising said compound. Insome embodiments, the bacteria infection is urinary tract infection orinflammatory bowel disease. In some embodiments, the bacteria infectionis ulcerative colitis. In other embodiments, the bacteria infection isCrohn's disease. In some embodiments, bacteria infection is the cause ofCrohn's Disease or ulcerative colitis. In some embodiments, the bacteriainfections are caused by AIEC (adherent-invasive E. coli) strains.

Another embodiment provides a method of treating or preventinginflammatory bowel disease in a subject, comprising administering to thesubject an effective amount of a compound described herein or apharmaceutically acceptable salt thereof, or a composition comprisingsaid compound. In some embodiments, the subject is a patient. In otherembodiments, the subject is a human. In some embodiments, theinflammatory bowel disease is Crohn's Disease. In other embodiments, theinflammatory bowel disease is ulcerative colitis.

Another embodiment provides a method of inhibiting FimH in bacteria froman E. coli bacterial strain isolated from patients with inflammatorybowel disease, comprising contacting the bacteria with an effectiveamount of a compound described herein, or a pharmaceutically acceptablesalt thereof, or a composition comprising said compound. In someembodiments, the bacterial strain is LF-82.

Another embodiment provides a method of inhibiting FimH in a subject,comprising administering to the subject an effective amount of acompound described herein, or a pharmaceutically acceptable saltthereof, or a composition comprising said compound.

Another embodiment provides a method of inhibiting adhesion of E. coliin a subject, comprising administering to the subject an effectiveamount of a compound described herein, or a pharmaceutically acceptablesalt thereof, or a composition comprising said compound. In someembodiments, the inhibition of adhesion results in the prevention of theestablishment of a sub-musosal infection.

Another embodiment provides a method of blocking the interaction betweentype 1 pili and CEACAM6 in a subject, comprising administering to thesubject an effective amount of a compound described herein, or apharmaceutically acceptable salt thereof, or a composition comprisingsaid compound.

As described herein, a specified number range of atoms includes anyinteger therein. For example, a group having from 1-4 atoms could have1, 2, 3, or 4 atoms.

The term “stable”, as used herein, refers to compounds that are notsubstantially altered when subjected to conditions to allow for theirproduction, detection, recovery, storage, purification, and use for oneor more of the purposes disclosed herein. In some embodiments, a stablecompound or chemically feasible compound is one that is notsubstantially altered when kept at a temperature of 40° C. or less, inthe absence of moisture or other chemically reactive conditions, for atleast a week.

The term “aliphatic” or “aliphatic group”, as used herein, means astraight-chain (i.e., unbranched), or branched, hydrocarbon chain thatis completely saturated or that contains one or more units ofunsaturation but is non-aromatic.

Unless otherwise specified, aliphatic groups contain 1-20 aliphaticcarbon atoms. In some embodiments, aliphatic groups contain 1-10aliphatic carbon atoms. In other embodiments, aliphatic groups contain1-8 aliphatic carbon atoms. In still other embodiments, aliphatic groupscontain 1-6 aliphatic carbon atoms, and in yet other embodimentsaliphatic groups contain 1-4 aliphatic carbon atoms. Aliphatic groupsmay be linear or branched, substituted or unsubstituted alkyl, alkenyl,or alkynyl groups. Specific examples include, but are not limited to,methyl, ethyl, isopropyl, n-propyl, sec-butyl, vinyl, n-butenyl,ethynyl, and tert-butyl. Alkyl groups can also include deuteratedhydrogens and include groups like CD₃.

The term “alkyl” as used herein means a saturated straight or branchedchain hydrocarbon. The term “alkenyl” as used herein means a straight orbranched chain hydrocarbon comprising one or more double bonds. The term“alkynyl” as used herein means a straight or branched chain hydrocarboncomprising one or more triple bonds. The term “cycloaliphatic” (or“carbocycle” or “carbocyclyl” or “carbocyclic”) refers to a non-aromaticmonocyclic carbon containing ring which can be saturated or contain oneor more units of unsaturation, having three to fourteen ring carbonatoms. The term includes polycyclic fused, spiro or bridged carbocyclicring systems. The term also includes polycyclic ring systems in whichthe carbocyclic ring can be fused to one or more non-aromaticcarbocyclic or heterocyclic rings or one or more aromatic rings orcombination thereof, wherein the radical or point of attachment is onthe carbocyclic ring. Fused bicyclic ring systems comprise two ringswhich share two adjoining ring atoms, bridged bicyclic group comprisetwo rings which share three or four adjacent ring atoms, spiro bicyclicring systems share one ring atom. Examples of cycloaliphatic groupsinclude, but are not limited to, cycloalkyl and cycloalkenyl groups.Specific examples include, but are not limited to, cyclohexyl,cyclopropenyl, and cyclobutyl. The term “heterocycle” (or“heterocyclyl”, or “heterocyclic”) as used herein means refers to anon-aromatic monocyclic ring which can be saturated or contain one ormore units of unsaturation, having three to fourteen ring atoms in whichone or more ring carbons is replaced by a heteroatom such as, N, S, orO. The term includes polycyclic fused, spiro or bridged heterocyclicring systems. The term also includes polycyclic ring systems in whichthe heterocyclic ring can be fused to one or more non-aromaticcarbocyclic or heterocyclic rings or one or more aromatic rings orcombination thereof, wherein the radical or point of attachment is onthe heterocyclic ring. Examples of heterocycles include, but are notlimited to, piperidinyl, piperizinyl, pyrrolidinyl, pyrazolidinyl,imidazolidinyl, azepanyl, diazepanyl, triazepanyl, azocanyl, diazocanyl,triazocanyl, oxazolidinyl, isoxazolidinyl, thiazolidinyl,isothiazolidinyl, oxazocanyl, oxazepanyl, thiazepanyl, thiazocanyl,benzimidazolonyl, tetrahydrofuranyl, tetrahydrofuranyl,tetrahydrothiophenyl, tetrahydrothiophenyl, morpholino, including, forexample, 3-morpholino, 4-morpholino, 2-thiomorpholino, 3-thiomorpholino,4-thiomorpholino, 1-pyrrolidinyl, 2-pyrrolidinyl, 3-pyrrolidinyl,1-tetrahydropiperazinyl, 2-tetrahydropiperazinyl,3-tetrahydropiperazinyl, 1-piperidinyl, 2-piperidinyl, 3-piperidinyl,1-pyrazolinyl, 3-pyrazolinyl, 4-pyrazolinyl, 5-pyrazolinyl,1-piperidinyl, 2-piperidinyl, 3-piperidinyl, 4-piperidinyl,2-thiazolidinyl, 3-thiazolidinyl, 4-thiazolidinyl, 1-imidazolidinyl,2-imidazolidinyl, 4-imidazolidinyl, 5-imidazolidinyl, indolinyl,tetrahydroquinolinyl, tetrahydroisoquinolinyl, thienothienyl,thienothiazolyl, benzothiolanyl, benzodithianyl,3-(1-alkyl)-benzimidazol-2-onyl, and 1,3-dihydro-imidazol-2-onyl.

The term “heteroatom” means one or more of oxygen, sulfur, nitrogen,phosphorus, or silicon (including, any oxidized form of nitrogen,sulfur, phosphorus, or silicon; the quaternized form of any basicnitrogen or; a substitutable nitrogen of a heterocyclic ring, forexample N (as in 3,4-dihydro-2H-pyrrolyl), NH (as in pyrrolidinyl) orNR⁺ (as in N-substituted pyrrolidinyl)).

The term “unsaturated”, as used herein, means that a moiety has one ormore units of unsaturation.

The term “alkoxy”, or “thioalkyl”, as used herein, refers to an alkylgroup, as previously defined, attached to the molecule through an oxygen(“alkoxy” e.g., —O-alkyl) or sulfur (“thioalkyl” e.g., —S-alkyl) atom.

The terms “haloalkyl”, “haloalkenyl”, “haloaliphatic”, and “haloalkoxy”mean alkyl, alkenyl or alkoxy, as the case may be, substituted with oneor more halogen atoms. This term includes perfluorinated alkyl groups,such as —CF₃ and —CF₂CF₃.

The terms “halogen”, “halo”, and “hal” mean F, Cl, Br, or I.

The term “aryl” used alone or as part of a larger moiety as in“aralkyl”, “aralkoxy”, or “aryloxyalkyl”, refers to carbocyclic aromaticring systems. The term “aryl” may be used interchangeably with the term“aryl ring”.

Carbocyclic aromatic ring groups have only carbon ring atoms (typicallysix to fourteen) and include monocyclic aromatic rings such as phenyland fused polycyclic aromatic ring systems in which two or morecarbocyclic aromatic rings are fused to one another. Examples include1-naphthyl, 2-naphthyl, 1-anthracyl and 2-anthracyl. Also includedwithin the scope of the term “carbocyclic aromatic ring”, as it is usedherein, is a group in which an aromatic ring is fused to one or morenon-aromatic rings (carbocyclic or heterocyclic), such as in an indanyl,phthalimidyl, naphthimidyl, phenanthridinyl, or tetrahydronaphthyl,where the radical or point of attachment is on the aromatic ring.

The term “heteroaryl”, “heteroaromatic”, “heteroaryl ring”, “heteroarylgroup” and “heteroaromatic group”, used alone or as part of a largermoiety as in “heteroaralkyl” or “heteroarylalkoxy”, refers toheteroaromatic ring groups having five to fourteen members, includingmonocyclic heteroaromatic rings and polycyclic aromatic rings in which amonocyclic aromatic ring is fused to one or more other aromatic ring.Heteroaryl groups have one or more ring heteroatoms. Also includedwithin the scope of the term “heteroaryl”, as it is used herein, is agroup in which an aromatic ring is fused to one or more non-aromaticrings (carbocyclic or heterocyclic), where the radical or point ofattachment is on the aromatic ring. Bicyclic 6,5 heteroaromatic ring, asused herein, for example, is a six membered heteroaromatic ring fused toa second five membered ring, wherein the radical or point of attachmentis on the six membered ring. Examples of heteroaryl groups includepyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, imidazolyl, pyrrolyl,pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl,thiazolyl, isothiazolyl or thiadiazolyl including, for example,2-furanyl, 3-furanyl, N-imidazolyl, 2-imidazolyl, 4-imidazolyl,5-imidazolyl, 3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl, 2-oxadiazolyl,5-oxadiazolyl, 2-oxazolyl, 4-oxazolyl, 5-oxazolyl, 3-pyrazolyl,4-pyrazolyl, 1-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl, 2-pyridyl, 3-pyridyl,4-pyridyl, 2-pyrimidinyl, 4-pyrimidinyl, 5-pyrimidinyl, 3-pyridazinyl,2-thiazolyl, 4-thiazolyl, 5-thiazolyl, 2-triazolyl, 5-triazolyl,tetrazolyl, 2-thienyl, 3-thienyl, carbazolyl, benzimidazolyl,benzothienyl, benzofuranyl, indolyl, benzotriazolyl, benzothiazolyl,benzoxazolyl, benzimidazolyl, isoquinolinyl, indolyl, isoindolyl,acridinyl, benzisoxazolyl, isothiazolyl, 1,2,3-oxadiazolyl,1,2,5-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,3-triazolyl,1,2,3-thiadiazolyl, 1,3,4-thiadiazolyl, 1,2,5-thiadiazolyl, purinyl,pyrazinyl, 1,3,5-triazinyl, quinolinyl (e.g., 2-quinolinyl,3-quinolinyl, 4-quinolinyl), and isoquinolinyl (e.g., 1-isoquinolinyl,3-isoquinolinyl, or 4-isoquinolinyl).

The term “protecting group” and “protective group” as used herein, areinterchangeable and refer to an agent used to temporarily block one ormore desired functional groups in a compound with multiple reactivesites. In certain embodiments, a protecting group has one or more, orpreferably all, of the following characteristics: a) is addedselectively to a functional group in good yield to give a protectedsubstrate that is b) stable to reactions occurring at one or more of theother reactive sites; and c) is selectively removable in good yield byreagents that do not attack the regenerated, deprotected functionalgroup. As would be understood by one skilled in the art, in some cases,the reagents do not attack other reactive groups in the compound. Inother cases, the reagents may also react with other reactive groups inthe compound. Examples of protecting groups are detailed in Greene, T.W., Wuts, P. G in “Protective Groups in Organic Synthesis”, ThirdEdition, John Wiley & Sons, New York: 1999 (and other editions of thebook), the entire contents of which are hereby incorporated byreference. The term “nitrogen protecting group”, as used herein, refersto an agent used to temporarily block one or more desired nitrogenreactive sites in a multifunctional compound. Preferred nitrogenprotecting groups also possess the characteristics exemplified for aprotecting group above, and certain exemplary nitrogen protecting groupsare also detailed in Chapter 7 in Greene, T. W., Wuts, P. G in“Protective Groups in Organic Synthesis”, Third Edition, John Wiley &Sons, New York: 1999, the entire contents of which are herebyincorporated by reference.

In some embodiments, where indicated a methylene unit of an aliphaticchain is optionally replaced with another atom or group. Examples ofsuch atoms or groups include, but are not limited to, —NR—, —O—, —C(O)—,—C(═N—CN)—, —C(═NR)—, —C(═NOR)—, —S—, —S(O)—, and —S(O)₂—. These atomsor groups can be combined to form larger groups. Examples of such largergroups include, but are not limited to, —OC(O)—, —C(O)CO—, —CO₂—,—C(O)NR—, —C(═N—CN), —NRC(O)—, —NRC(O)O—, —S(O)₂NR—, —NRSO₂—,—NRC(O)NR—, —OC(O)NR—, and —NRSO₂NR—, wherein R is defined herein.

Only those replacement and combinations of groups that result in astable structure are contemplated. Optional replacements can occur bothwithin the chain and/or at either end of the chain; i.e. both at thepoint of attachment and/or also at the terminal end. Two optionalreplacements can also be adjacent to each other within a chain so longas it results in a chemically stable compound. The optional replacementscan also completely replace all of the carbon atoms in a chain. Forexample, a C₃ aliphatic can be optionally replaced by —NR—, —C(O)—, and—NR— to form —NRC(O)NR— (a urea).

Unless otherwise indicated, if the replacement occurs at the terminalend, the replacement atom is bound to an H on the terminal end. Forexample, if —CH₂CH₂CH₃ were optionally replaced with —O—, the resultingcompound could be —OCH₂CH₃, —CH₂OCH₃, or —CH₂CH₂OH.

It shall be understood that an aliphatic chain may include bonds ofunsaturation, and therefore the atom which is replacing the “methylene”unit of an aliphatic may in fact be replacing a —CH═ unit, a ═C═ unit ora ≡C— unit. One of skill in the art would understand that the atomreplacing these units would have the appropriate bond order to result ina stable structure. For example, when a methylene unit of an aliphaticchain is optionally replaced with —NR—, one of skill in the art wouldunderstand that if the aliphatic group were CH═CH—CH₃ and the middlemethylene group, “CH” were to be replaced, it would actually be replacedwith “N”, not “—NR—” to result in CH═N—CH₃.

Unless otherwise indicated, structures depicted herein are also meant toinclude all isomeric (e.g., enantiomeric, diastereomeric, geometric,conformational, and rotational) forms of the structure. For example, theR and S configurations for each asymmetric center, (Z) and (E) doublebond isomers, and (Z) and (E) conformational isomers are included inthis invention. As would be understood to one skilled in the art, asubstituent can freely rotate around any rotatable bonds. For example, asubstituent drawn as

also represents

Therefore, single stereochemical isomers as well as enantiomeric,diastereomeric, geometric, conformational, and rotational mixtures ofthe present compounds are within the scope of the invention.

Unless otherwise indicated, all tautomeric forms of the compounds of theinvention are within the scope of the invention.

Additionally, unless otherwise indicated, structures depicted herein arealso meant to include compounds that differ only in the presence of oneor more isotopically enriched atoms. For example, compounds having thepresent structures except for the replacement of hydrogen by deuteriumor tritium, or the replacement of a carbon by a ¹³C- or ¹⁴C-enrichedcarbon are within the scope of this invention. Such compounds areuseful, for example, as analytical tools or probes in biological assays.These compounds are also useful in the treatment of bacteria infections(see e.g., compound 183).

As described herein, where indicated compounds of the invention mayoptionally be substituted with one or more substituents, such as areillustrated generally herein, or as exemplified by particular classes,subclasses, and species of the invention. It will be appreciated thatthe phrase “optionally substituted” is used interchangeably with thephrase “substituted or unsubstituted.” In general, the term“substituted”, whether preceded by the term “optionally” or not, refersto the replacement of hydrogen radicals in a given structure with theradical of a specified substituent. Unless otherwise indicated, anoptionally substituted group may have a substituent at eachsubstitutable position of the group, and when more than one position inany given structure may be substituted with more than one substituentselected from a specified group, the substituent may be either the sameor different at every position.

Only those choices and combinations of substituents that result in astable structure are contemplated. Such choices and combinations will beapparent to those of ordinary skill in the art and may be determinedwithout undue experimentation.

The term “ring atom” is an atom such as C, N, O or S that is in the ringof an aromatic group, cycloalkyl group or non-aromatic heterocyclicring.

A “substitutable ring atom” in an aromatic group is a ring carbon ornitrogen atom bonded to a hydrogen atom. The hydrogen can be optionallyreplaced with a suitable substituent group. Thus, the term“substitutable ring atom” does not include ring nitrogen or carbon atomswhich are shared when two rings are fused. In addition, “substitutablering atom” does not include ring carbon or nitrogen atoms when thestructure depicts that they are already attached to a moiety other thanhydrogen.

It shall be understood that a bond with a substituent drawn throughseveral rings of a polycyclic molecule indicates that the substituentmay be bonded at any ring of the polycyclic ring. For example, in thefigure shown below:

J^(H) can be substituted on either benzo ring of the carbazolyl ring, aswell as on the 5-membered ring in the center, such as at the nitrogen ofthe carbazolyl ring. Therefore, if J^(H) were “CH₃”, any of thefollowing three would be comtemplated based on the above formula:

An aryl group as defined herein may contain one or more substitutablering atoms, which may be bonded to a suitable substituent. Examples ofsuitable substituents on a substitutable ring carbon atom of an arylgroup include R′. R′ is —Ra, —Br, —Cl, —I, —F, —ORa, —SRa, —O—CORa,—CORa, —CSRa, —CN, —NO₂, —NCS, —SO₃H, —N(RaRb), —COORa, —NRcNRcCORa,—NRcNRcCO₂Ra, —CHO, —CON(RaRb), —OC(O)N(RaRb), —CSN(RaRb), —NRcCORa,—NRcCOORa, —NRcCSRa, —NRcCON(RaRb), —NRcNRcC(O)N(RaRb), —NRcCSN(RaRb),—C(═NRc)-N(RaRb), —C(═S)N(RaRb), —NRd-C(═NRc)-N(RaRb), —NRcNRaRb,—S(O)_(p)NRaRb, —NRcSO₂N(RaRb), —NRcS(O)_(p)Ra, —S(O)_(p)Ra,—OS(O)_(p)NRaRb or —OS(O)_(p)Ra; wherein p is 1 or 2.

Ra-Rd are each independently —H, an aliphatic group, aromatic group,non-aromatic carbocyclic or heterocyclic group or —N(RaRb), takentogether, form a non-aromatic heterocyclic group. The aliphatic,aromatic and non-aromatic heterocyclic group represented by Ra-Rd andthe non-aromatic heterocyclic group represented by —N(RaRb) are eachoptionally and independently substituted with one or more groupsrepresented by R^(#). Preferably Ra-Rd are unsubstituted.

R^(#) is halogen, R⁺, —OR⁺, —SR⁺, —NO₂, —CN, —N(R⁺)₂, —COR⁺, —COOR⁺,—NHCO₂R⁺, —NHC(O)R⁺, —NHNHC(O)R⁺, —NHC(O)N(R⁺)₂, —NHNHC(O)N(R⁺)₂,—NHNHCO₂R⁺, —C(O)N(R⁺)₂, —OC(O)R⁺, —OC(O)N(R⁺)₂, —S(O)₂R⁺, —SO₂N(R⁺)₂,—S(O)R⁺, —NHSO₂N(R⁺)₂, —NHSO₂R⁺, —C(═S)N(R⁺)₂, or —C(═NH)—N(R⁺)₂.

R⁺ is —H, a C1-C4 alkyl group, a monocyclic aryl group, a non-aromaticcarbocyclic or heterocyclic group each optionally substituted withalkyl, haloalkyl, alkoxy, haloalkoxy, halo, —CN, —NO₂, amine, alkylamineor dialkylamine. Preferably R+ is unsubstituted.

An aliphatic or a non-aromatic heterocyclic or carbocyclic group as usedherein may contain one or more substituents. Examples of suitablesubstituents for an aliphatic group or a ring carbon of a non-aromaticheterocyclic group is R″. R″ include those substituents listed above forR′ and ═O, ═S, ═NNHR**, ═NN(R**)2, ═NNHC(O)R**, ═NNHCO2 (alkyl), ═NNHSO2(alkyl), ═NR**, spiro cycloalkyl group or fused cycloalkyl group. EachR** is independently selected from hydrogen, an unsubstituted alkylgroup or a substituted alkyl group. Examples of substituents on thealkyl group represented by R** include amino, alkylamino, dialkylamino,aminocarbonyl, halogen, alkyl, alkylaminocarbonyl, dialkylaminocarbonyl,alkylaminocarbonyloxy, dialkylaminocarbonyloxy, alkoxy, nitro, cyano,carboxy, alkoxycarbonyl, alkylcarbonyl, hydroxy, haloalkoxy, orhaloalkyl.

When a heterocyclyl, heteroaryl, or heteroaralkyl group contains anitrogen atom, it may be substituted or unsubstituted. When a nitrogenatom in the aromatic ring of a heteroaryl group has a substituent thenitrogen may be a quaternary nitrogen.

A preferred position for substitution of a non-aromaticnitrogen-containing heterocyclic group is the nitrogen ring atom.Suitable substituents on the nitrogen of a non-aromatic heterocyclicgroup or heteroaryl group include —R{circumflex over ( )},—N(R{circumflex over ( )})₂, C(O)R{circumflex over ( )}, CO₂R{circumflexover ( )}, —C(O)C(O)R{circumflex over ( )}, —SO₂R{circumflex over ( )},SO₂N(R{circumflex over ( )})₂, C(═S)N(R{circumflex over ( )})₂,C(═NH)—N(R{circumflex over ( )})₂, and —NR{circumflex over( )}SO₂R{circumflex over ( )}; wherein R{circumflex over ( )} ishydrogen, an aliphatic group, a substituted aliphatic group, aryl,substituted aryl, heterocyclic or carbocyclic ring or a substitutedheterocyclic or carbocyclic ring. Examples of substituents on the grouprepresented by R{circumflex over ( )} include alkyl, haloalkoxy,haloalkyl, alkoxyalkyl, sulfonyl, alkylsulfonyl, halogen, nitro, cyano,hydroxy, aryl, carbocyclic or heterocyclic ring, oxo, amino, alkylamino,dialkylamino, aminocarbonyl, alkylaminocarbonyl,dialkylaminocarbonyloxy, alkoxy, carboxy, alkoxycarbonyl, oralkylcarbonyl. Preferably R{circumflex over ( )} is not substituted.

Non-aromatic nitrogen containing heterocyclic rings that are substitutedon a ring nitrogen and attached to the remainder of the molecule at aring carbon atom are said to be N substituted. For example, an N alkylpiperidinyl group is attached to the remainder of the molecule at thetwo, three or four position of the piperidinyl ring and substituted atthe ring nitrogen with an alkyl group. Non-aromatic nitrogen containingheterocyclic rings such as pyrazinyl that are substituted on a ringnitrogen and attached to the remainder of the molecule at a second ringnitrogen atom are said to be N′ substituted-N-heterocycles. For example,an N′ acyl N-pyrazinyl group is attached to the remainder of themolecule at one ring nitrogen atom and substituted at the second ringnitrogen atom with an acyl group.

As used herein an optionally substituted aralkyl can be substituted onboth the alkyl and the aryl portion. Unless otherwise indicated as usedherein optionally substituted aralkyl is optionally substituted on thearyl portion.

The terms “a bond” and “absent” are used interchangeably to indicatethat a group is absent.

The compounds of the invention are defined herein by their chemicalstructures and/or chemical names. Where a compound is referred to byboth a chemical structure and a chemical name, and the chemicalstructure and chemical name conflict, the chemical structure isdeterminative of the compound's identity.

The compounds of this invention can exist in free form for treatment, orwhere appropriate, as a pharmaceutically acceptable salt.

As used herein, the term “pharmaceutically acceptable salt” refers tosalts of a compound which are, within the scope of sound medicaljudgment, suitable for use in contact with the tissues of humans andlower animals without undue side effects, such as, toxicity, irritation,allergic response and the like, and are commensurate with a reasonablebenefit/risk ratio.

Pharmaceutically acceptable salts are well known in the art. Forexample, S. M. Berge et al., describe pharmaceutically acceptable saltsin detail in J. Pharmaceutical Sciences, 1977, 66, 1-19, incorporatedherein by reference. Pharmaceutically acceptable salts of the compoundsof this invention include those derived from suitable inorganic andorganic acids and bases. These salts can be prepared in situ during thefinal isolation and purification of the compounds. Acid addition saltscan be prepared by 1) reacting the purified compound in its free-basedform with a suitable organic or inorganic acid and 2) isolating the saltthus formed.

Examples of pharmaceutically acceptable, nontoxic acid addition saltsare salts of an amino group formed with inorganic acids such ashydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid andperchloric acid or with organic acids such as acetic acid, oxalic acid,maleic acid, tartaric acid, citric acid, succinic acid or malonic acidor by using other methods used in the art such as ion exchange. Otherpharmaceutically acceptable salts include adipate, alginate, ascorbate,aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate,camphorate, camphorsulfonate, citrate, cyclopentanepropionate,digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate,glucoheptonate, glycerophosphate, glycolate, gluconate, glycolate,hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide,hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate,lauryl sulfate, malate, maleate, malonate, methanesulfonate,2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate,palmoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate,pivalate, propionate, salicylate, stearate, succinate, sulfate,tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate salts,and the like.

Base addition salts can be prepared by 1) reacting the purified compoundin its acid form with a suitable organic or inorganic base and 2)isolating the salt thus formed. Salts derived from appropriate basesinclude alkali metal (e.g., sodium, lithium, and potassium), alkalineearth metal (e.g., magnesium and calcium), ammonium and N⁺(C₁₋₄alkyl)₄salts. This invention also envisions the quaternization of any basicnitrogen-containing groups of the compounds disclosed herein. Water oroil-soluble or dispersible products may be obtained by suchquaternization.

Further pharmaceutically acceptable salts include, when appropriate,nontoxic ammonium, quaternary ammonium, and amine cations formed usingcounterions such as halide, hydroxide, carboxylate, sulfate, phosphate,nitrate, lower alkyl sulfonate and aryl sulfonate. Other acids andbases, while not in themselves pharmaceutically acceptable, may beemployed in the preparation of salts useful as intermediates inobtaining the compounds of the invention and their pharmaceuticallyacceptable acid or base addition salts.

It should be understood that this invention includesmixtures/combinations of different pharmaceutically acceptable salts andalso mixtures/combinations of compounds in free form andpharmaceutically acceptable salts.

In addition to the compounds of this invention, pharmaceuticallyacceptable derivatives or prodrugs of the compounds of this inventionmay also be employed in compositions to treat or prevent the hereinidentified disorders.

As used herein and unless otherwise indicated, the term “prodrug” meansa derivative of a compound that can hydrolyze, oxidize, or otherwisereact under biological conditions (in vitro or in vivo) to provide acompound of this invention. Prodrugs may become active upon suchreaction under biological conditions, or they may have activity in theirunreacted forms. Examples of prodrugs contemplated in this inventioninclude, but are not limited to, analogs or derivatives of compounds ofthe invention that comprise biohydrolyzable moieties such asbiohydrolyzable amides, biohydrolyzable esters, biohydrolyzablecarbamates, biohydrolyzable carbonates, biohydrolyzable ureides, andbiohydrolyzable phosphate analogues. Other examples of prodrugs includederivatives of compounds of the invention that comprise —NO, —NO₂, —ONO,or —ONO₂ moieties. Prodrugs can typically be prepared using well-knownmethods, such as those described by BURGER'S MEDICINAL CHEMISTRY ANDDRUG DISCOVERY (1995) 172-178, 949-982 (Manfred E. Wolff ed., 5th ed).

A “pharmaceutically acceptable derivative” is an adduct or derivativewhich, upon administration to a patient in need, is capable ofproviding, directly or indirectly, a compound as otherwise describedherein, or a metabolite or residue thereof. Examples of pharmaceuticallyacceptable derivatives include, but are not limited to, esters and saltsof such esters.

A “pharmaceutically acceptable derivative or prodrug” includes anypharmaceutically acceptable ester, salt of an ester or other derivativeor salt thereof of a compound, of this invention which, uponadministration to a recipient, is capable of providing, either directlyor indirectly, a compound of this invention or an inhibitorily activemetabolite or residue thereof. Particularly favoured derivatives orprodrugs are those that increase the bioavailability of the compounds ofthis invention when such compounds are administered to a patient (e.g.,by allowing an orally administered compound to be more readily absorbedinto the blood) or which enhance delivery of the parent compound to abiological compartment (e.g., the brain or lymphatic system) relative tothe parent species.

Pharmaceutically acceptable prodrugs of the compounds of this inventioninclude, without limitation, esters, amino acid esters, phosphateesters, metal salts and sulfonate esters.

As used herein, the phrase “side effects” encompasses unwanted andadverse effects of a therapy (e.g., a prophylactic or therapeuticagent). Side effects are always unwanted, but unwanted effects are notnecessarily adverse. An adverse effect from a therapy (e.g.,prophylactic or therapeutic agent) might be harmful or uncomfortable orrisky. Side effects include, but are not limited to fever, chills,lethargy, gastrointestinal toxicities (including gastric and intestinalulcerations and erosions), nausea, vomiting, neurotoxicities,nephrotoxicities, renal toxicities (including such conditions aspapillary necrosis and chronic interstitial nephritis), hepatictoxicities (including elevated serum liver enzyme levels),myelotoxicities (including leukopenia, myelosuppression,thrombocytopenia and anemia), dry mouth, metallic taste, prolongation ofgestation, weakness, somnolence, pain (including muscle pain, bone painand headache), hair loss, asthenia, dizziness, extra-pyramidal symptoms,akathisia, cardiovascular disturbances and sexual dysfunction.

In one embodiment the present invention is a pharmaceutical compositioncomprising a compound of the present invention and a pharmaceuticallyacceptable carrier, diluent, adjuvant or vehicle. In one embodiment thepresent invention is a pharmaceutical composition comprising aneffective amount of compound of the present invention and apharmaceutically acceptable carrier, diluent, adjuvant or vehicle.Pharmaceutically acceptable carriers include, for example,pharmaceutical diluents, excipients or carriers suitably selected withrespect to the intended form of administration, and consistent withconventional pharmaceutical practices.

A pharmaceutically acceptable carrier may contain inert ingredientswhich do not unduly inhibit the biological activity of the compounds.The pharmaceutically acceptable carriers should be biocompatible, e.g.,non-toxic, non-inflammatory, non-immunogenic or devoid of otherundesired reactions or side-effects upon the administration to asubject. Standard pharmaceutical formulation techniques can be employed.

The pharmaceutically acceptable carrier, adjuvant, or vehicle, as usedherein, includes any and all solvents, diluents, or other liquidvehicle, dispersion or suspension aids, surface active agents, isotonicagents, thickening or emulsifying agents, preservatives, solid binders,lubricants and the like, as suited to the particular dosage formdesired. Remington's Pharmaceutical Sciences, Sixteenth Edition, E. W.Martin (Mack Publishing Co., Easton, Pa., 1980) discloses variouscarriers used in formulating pharmaceutically acceptable compositionsand known techniques for the preparation thereof. Except insofar as anyconventional carrier medium is incompatible with the compounds of theinvention, such as by producing any undesirable biological effect orotherwise interacting in a deleterious manner with any othercomponent(s) of the pharmaceutically acceptable composition, its use iscontemplated to be within the scope of this invention.

Some examples of materials which can serve as pharmaceuticallyacceptable carriers include, but are not limited to, ion exchangers,alumina, aluminum stearate, lecithin, serum proteins, such as humanserum albumin, buffer substances such as phosphates, glycine, sorbicacid, or potassium sorbate, partial glyceride mixtures of saturatedvegetable fatty acids, water, salts or electrolytes, such as protaminesulfate, disodium hydrogen phosphate, potassium hydrogen phosphate,sodium chloride, zinc salts, colloidal silica, magnesium trisilicate,polyvinyl pyrrolidone, polyacrylates, waxes,polyethylene-polyoxypropylene-block polymers, wool fat, sugars such aslactose, glucose and sucrose; starches such as corn starch and potatostarch; cellulose and its derivatives such as sodium carboxymethylcellulose, ethyl cellulose and cellulose acetate; powdered tragacanth;malt; gelatin; talc; excipients such as cocoa butter and suppositorywaxes; oils such as peanut oil, cottonseed oil; safflower oil; sesameoil; olive oil; corn oil and soybean oil; glycols; such a propyleneglycol or polyethylene glycol; esters such as ethyl oleate and ethyllaurate; agar; buffering agents such as magnesium hydroxide and aluminumhydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer'ssolution; ethyl alcohol, and phosphate buffer solutions, as well asother non-toxic compatible lubricants such as sodium lauryl sulfate andmagnesium stearate, as well as coloring agents, releasing agents,coating agents, sweetening, flavoring and perfuming agents,preservatives and antioxidants can also be present in the composition,according to the judgment of the formulator.

The compounds of present invention or pharmaceutical salts thereof maybe formulated into pharmaceutical compositions for administration to asubject as defined herein. These pharmaceutical compositions, whichcomprise an amount of the compounds effective to treat or prevent abacteria infection, such as IBD, and a pharmaceutically acceptablecarrier, are another embodiment of the present invention.

In one embodiment the present invention is a method of treating orpreventing a bacteria infection, such as IBD, in a subject in needthereof, comprising administering to the subject an effective amount ofa compound or composition of the present invention.

As used herein, the terms “subject”, “patient” and “mammal” are usedinterchangeably. The terms “subject” and “patient” refer to an animal(e.g., a bird such as a chicken, quail or turkey, or a mammal),preferably a mammal including a non-primate (e.g., a cow, pig, horse,sheep, rabbit, guinea pig, rat, cat, dog, and mouse) and a primate(e.g., a monkey, chimpanzee and a human), and more preferably a human.In one embodiment, the subject is a non-human animal such as a farmanimal (e.g., a horse, cow, pig or sheep), or a pet (e.g., a dog, cat,guinea pig or rabbit). In a preferred embodiment, the subject is ahuman.

As used herein, an “effective amount” refers to an amount sufficient toelicit the desired biological response. In the present invention thedesired biological response is to reduce or ameliorate the severity,duration, progression, or onset of a bacteria infection, prevent theadvancement of a bacteria infection, cause the regression of a bacteriainfection, prevent the recurrence, development, onset or progression ofa symptom associated with a bacteria infection, or enhance or improvethe prophylactic or therapeutic effect(s) of another therapy. Theprecise amount of compound administered to a subject will depend on themode of administration, the type and severity of the disease orcondition and on the characteristics of the subject, such as generalhealth, age, sex, body weight and tolerance to drugs. It will alsodepend on the degree, severity and type of bacteria infection, and themode of administration. The skilled artisan will be able to determineappropriate dosages depending on these and other factors. Whenco-administered with other agents, e.g., when co-administered with abacteria infection agent, an “effective amount” of the second agent willdepend on the type of drug used. Suitable dosages are known for approvedagents and can be adjusted by the skilled artisan according to thecondition of the subject, the type of condition(s) being treated and theamount of a compound of the invention being used. In cases where noamount is expressly noted, an effective amount should be assumed.

As used herein, the terms “treat”, “treatment” and “treating” refer tothe reduction or amelioration of the progression, severity and/orduration of a bacteria infection, or the amelioration of one or moresymptoms (preferably, one or more discernible symptoms) of a bacteriainfection resulting from the administration of one or more therapies(e.g., one or more therapeutic agents such as a compound of theinvention). In specific embodiments, the terms “treat”, “treatment” and“treating” refer to the amelioration of at least one measurable physicalparameter of a bacteria infection. In other embodiments the terms“treat”, “treatment” and “treating” refer to the inhibition of theprogression of a bacteria infection, either physically by, e.g.,stabilization of a discernible symptom, physiologically by, e.g.,stabilization of a physical parameter, or both. In other embodiments theterms “treat”, “treatment” and “treating” refer to the reduction orstabilization of a bacteria infection.

As used herein, the terms “prevent”, “prevention” and “preventing” referto the reduction in the risk of acquiring or developing a given bacteriainfection, or the reduction or inhibition of the recurrence or abacteria infection. In one embodiment, a compound of the invention isadministered as a preventative measure to a patient, preferably a human,having a genetic predisposition to any of the conditions, diseases ordisorders described herein.

The pharmaceutically acceptable compositions of this invention can beadministered to humans and other animals orally, rectally, parenterally,intracisternally, intravaginally, intraperitoneally, topically (as bypowders, ointments, or drops), bucally, as an oral or nasal spray, orthe like, depending on the severity of the infection being treated.Liquid dosage forms for oral administration include, but are not limitedto, pharmaceutically acceptable emulsions, microemulsions, solutions,suspensions, syrups and elixirs. In addition to the active compounds,the liquid dosage forms may contain inert diluents commonly used in theart such as, for example, water or other solvents, solubilizing agentsand emulsifiers such as ethyl alcohol, isopropyl alcohol, ethylcarbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propyleneglycol, 1,3-butylene glycol, dimethylformamide, oils (in particular,cottonseed, groundnut, corn, germ, olive, castor, and sesame oils),glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fattyacid esters of sorbitan, and mixtures thereof. Besides inert diluents,the oral compositions can also include adjuvants such as wetting agents,emulsifying and suspending agents, sweetening, flavoring, and perfumingagents.

Injectable preparations, for example, sterile injectable aqueous oroleaginous suspensions may be formulated according to the known artusing suitable dispersing or wetting agents and suspending agents. Thesterile injectable preparation may also be a sterile injectablesolution, suspension or emulsion in a nontoxic parenterally acceptablediluent or solvent, for example, as a solution in 1,3-butanediol. Amongthe acceptable vehicles and solvents that may be employed are water,Ringer's solution, U.S.P. and isotonic sodium chloride solution. Inaddition, sterile, fixed oils are conventionally employed as a solventor suspending medium. For this purpose any bland fixed oil can beemployed including synthetic mono- or diglycerides. In addition, fattyacids such as oleic acid are used in the preparation of injectables.

The injectable formulations can be sterilized, for example, byfiltration through a bacterial-retaining filter, or by incorporatingsterilizing agents in the form of sterile solid compositions which canbe dissolved or dispersed in sterile water or other sterile injectablemedium prior to use.

In order to prolong the effect of a compound of the present invention,it is often desirable to slow the absorption of the compound fromsubcutaneous or intramuscular injection. This may be accomplished by theuse of a liquid suspension of crystalline or amorphous material withpoor water solubility. The rate of absorption of the compound thendepends upon its rate of dissolution that, in turn, may depend uponcrystal size and crystalline form. Alternatively, delayed absorption ofa parenterally administered compound form is accomplished by dissolvingor suspending the compound in an oil vehicle. Injectable depot forms aremade by forming microencapsule matrices of the compound in biodegradablepolymers such as polylactide-polyglycolide. Depending upon the ratio ofcompound to polymer and the nature of the particular polymer employed,the rate of compound release can be controlled. Examples of otherbiodegradable polymers include poly(orthoesters) and poly(anhydrides).Depot injectable formulations are also prepared by entrapping thecompound in liposomes or microemulsions that are compatible with bodytissues.

Compositions for rectal or vaginal administration are preferablysuppositories which can be prepared by mixing the compounds of thisinvention with suitable non-irritating excipients or carriers such ascocoa butter, polyethylene glycol or a suppository wax which are solidat ambient temperature but liquid at body temperature and therefore meltin the rectum or vaginal cavity and release the active compound.

Solid dosage forms for oral administration include capsules, tablets,pills, powders, and granules. In such solid dosage forms, the activecompound is mixed with at least one inert, pharmaceutically acceptableexcipient or carrier such as sodium citrate or dicalcium phosphateand/or a) fillers or extenders such as starches, lactose, sucrose,glucose, mannitol, and silicic acid, b) binders such as, for example,carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone,sucrose, and acacia, c) humectants such as glycerol, d) disintegratingagents such as agar-agar, calcium carbonate, potato or tapioca starch,alginic acid, certain silicates, and sodium carbonate, e) solutionretarding agents such as paraffin, f) absorption accelerators such asquaternary ammonium compounds, g) wetting agents such as, for example,cetyl alcohol and glycerol monostearate, h) absorbents such as kaolinand bentonite clay, and i) lubricants such as talc, calcium stearate,magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate,and mixtures thereof. In the case of capsules, tablets and pills, thedosage form may also comprise buffering agents.

Solid compositions of a similar type may also be employed as fillers insoft and hard-filled gelatin capsules using such excipients as lactoseor milk sugar as well as high molecular weight polyethylene glycols andthe like. The solid dosage forms of tablets, dragees, capsules, pills,and granules can be prepared with coatings and shells such as entericcoatings and other coatings well known in the pharmaceutical formulatingart. They may optionally contain opacifying agents and can also be of acomposition that they release the active ingredient(s) only, orpreferentially, in a certain part of the intestinal tract, optionally,in a delayed manner. Examples of embedding compositions that can be usedinclude polymeric substances and waxes. Solid compositions of a similartype may also be employed as fillers in soft and hard-filled gelatincapsules using such excipients as lactose or milk sugar as well as highmolecular weight polyethylene glycols and the like.

The active compounds can also be in microencapsulated form with one ormore excipients as noted above. The solid dosage forms of tablets,dragees, capsules, pills, and granules can be prepared with coatings andshells such as enteric coatings, release controlling coatings and othercoatings well known in the pharmaceutical formulating art. In such soliddosage forms the active compound may be admixed with at least one inertdiluent such as sucrose, lactose or starch. Such dosage forms may alsocomprise, as is normal practice, additional substances other than inertdiluents, e.g., tableting lubricants and other tableting aids such amagnesium stearate and microcrystalline cellulose. In the case ofcapsules, tablets and pills, the dosage forms may also comprisebuffering agents. They may optionally contain opacifying agents and canalso be of a composition that they release the active ingredient(s)only, or preferentially, in a certain part of the intestinal tract,optionally, in a delayed manner. Examples of embedding compositions thatcan be used include polymeric substances and waxes.

Dosage forms for topical or transdermal administration of a compound ofthis invention include ointments, pastes, creams, lotions, gels,powders, solutions, sprays, inhalants or patches. The active componentis admixed under sterile conditions with a pharmaceutically acceptablecarrier and any needed preservatives or buffers as may be required.Ophthalmic formulation, eardrops, and eye drops are also contemplated asbeing within the scope of this invention. Additionally, the presentinvention contemplates the use of transdermal patches, which have theadded advantage of providing controlled delivery of a compound to thebody. Such dosage forms can be made by dissolving or dispensing thecompound in the proper medium. Absorption enhancers can also be used toincrease the flux of the compound across the skin. The rate can becontrolled by either providing a rate controlling membrane or bydispersing the compound in a polymer matrix or gel.

The compositions of the present invention may be administered orally,parenterally, by inhalation spray, topically, rectally, nasally,buccally, vaginally or via an implanted reservoir. The term “parenteral”as used herein includes, but is not limited to, subcutaneous,intravenous, intramuscular, intra-articular, intra-synovial,intrasternal, intrathecal, intrahepatic, intralesional and intracranialinjection or infusion techniques. Preferably, the compositions areadministered orally, intraperitoneally or intravenously.

Sterile injectable forms of the compositions of this invention may beaqueous or oleaginous suspension. These suspensions may be formulatedaccording to techniques known in the art using suitable dispersing orwetting agents and suspending agents. The sterile injectable preparationmay also be a sterile injectable solution or suspension in a non-toxicparenterally-acceptable diluent or solvent, for example as a solution in1,3-butanediol. Among the acceptable vehicles and solvents that may beemployed are water, Ringer's solution and isotonic sodium chloridesolution. In addition, sterile, fixed oils are conventionally employedas a solvent or suspending medium. For this purpose, any bland fixed oilmay be employed including synthetic mono- or di-glycerides. Fatty acids,such as oleic acid and its glyceride derivatives are useful in thepreparation of injectables, as are natural pharmaceutically-acceptableoils, such as olive oil or castor oil, especially in theirpolyoxyethylated versions. These oil solutions or suspensions may alsocontain a long-chain alcohol diluent or dispersant, such ascarboxymethyl cellulose or similar dispersing agents which are commonlyused in the formulation of pharmaceutically acceptable dosage formsincluding emulsions and suspensions. Other commonly used surfactants,such as Tweens, Spans and other emulsifying agents or bioavailabilityenhancers which are commonly used in the manufacture of pharmaceuticallyacceptable solid, liquid, or other dosage forms may also be used for thepurposes of formulation.

The pharmaceutical compositions of this invention may be orallyadministered in any orally acceptable dosage form including, but notlimited to, capsules, tablets, aqueous suspensions or solutions. In thecase of tablets for oral use, carriers commonly used include, but arenot limited to, lactose and corn starch. Lubricating agents, such asmagnesium stearate, are also typically added. For oral administration ina capsule form, useful diluents include lactose and dried cornstarch.When aqueous suspensions are required for oral use, the activeingredient is combined with emulsifying and suspending agents. Ifdesired, certain sweetening, flavoring or coloring agents may also beadded.

Alternatively, the pharmaceutical compositions of this invention may beadministered in the form of suppositories for rectal administration.These can be prepared by mixing the agent with a suitable non-irritatingexcipient which is solid at room temperature but liquid at rectaltemperature and therefore will melt in the rectum to release the drug.Such materials include, but are not limited to, cocoa butter, beeswaxand polyethylene glycols.The pharmaceutical compositions of this invention may also beadministered topically, especially when the target of treatment includesareas or organs readily accessible by topical application, includingdiseases of the eye, the skin, or the lower intestinal tract. Suitabletopical formulations are readily prepared for each of these areas ororgans.Topical application for the lower intestinal tract can be effected in arectal suppository formulation (see above) or in a suitable enemaformulation. Topically-transdermal patches may also be used.

For topical applications, the pharmaceutical compositions may beformulated in a suitable ointment containing the active componentsuspended or dissolved in one or more carriers. Carriers for topicaladministration of the compounds of this invention include, but are notlimited to, mineral oil, liquid petrolatum, white petrolatum, propyleneglycol, polyoxyethylene, polyoxypropylene compound, emulsifying wax andwater. Alternatively, the pharmaceutical compositions can be formulatedin a suitable lotion or cream containing the active components suspendedor dissolved in one or more pharmaceutically acceptable carriers.Suitable carriers include, but are not limited to, mineral oil, sorbitanmonostearate, polysorbate 60, cetyl esters wax, cetearyl alcohol, 2octyldodecanol, benzyl alcohol and water.

For ophthalmic use, the pharmaceutical compositions may be formulated asmicronized suspensions in isotonic, pH adjusted sterile saline, or,preferably, as solutions in isotonic, pH adjusted sterile saline, eitherwith or without a preservative such as benzylalkonium chloride.Alternatively, for ophthalmic uses, the pharmaceutical compositions maybe formulated in an ointment such as petrolatum.

The pharmaceutical compositions of this invention may also beadministered by nasal aerosol or inhalation. Such compositions areprepared according to techniques well-known in the art of pharmaceuticalformulation and may be prepared as solutions in saline, employing benzylalcohol or other suitable preservatives, absorption promoters to enhancebioavailability, fluorocarbons, and/or other conventional solubilizingor dispersing agents. The dosage regimen utilizing the compounds ofpresent invention can be selected in accordance with a variety offactors including the disorder being treated and the severity of thedisorder; the activity of the specific compound employed; the specificcomposition employed; the age, body weight, general health, sex and dietof the patient; the time of administration, route of administration, andrate of excretion of the specific compound employed; the renal andhepatic function of the subject; and the particular compound or saltthereof employed, the duration of the treatment; drugs used incombination or coincidental with the specific compound employed, andlike factors well known in the medical arts. The skilled artisan canreadily determine and prescribe the effective amount of the compound ofpresent invention required to treat, for example, to prevent, inhibit(fully or partially) or arrest the progress of the disease.

Dosages of the compounds of present invention can range from betweenabout 0.01 to about 100 mg/kg body weight/day, about 0.01 to about 50mg/kg body weight/day, about 0.1 to about 50 mg/kg body weight/day, orabout 1 to about 25 mg/kg body weight/day. It is understood that thetotal amount per day can be administered in a single dose or can beadministered in multiple dosings such as twice, three or four times perday.

The compounds for use in the method of the invention can be formulatedin unit dosage form. The term “unit dosage form” refers to physicallydiscrete units suitable as unitary dosage for subjects undergoingtreatment, with each unit containing a predetermined quantity of activematerial calculated to produce the desired therapeutic effect,optionally in association with a suitable pharmaceutical carrier. Theunit dosage form can be for a single daily dose or one of multiple dailydoses (e.g., about 1 to 4 or more times per day). When multiple dailydoses are used, the unit dosage form can be the same or different foreach dose.

An effective amount can be achieved in the method or pharmaceuticalcomposition of the invention employing a compound of present inventionor a pharmaceutically acceptable salt thereof alone or in combinationwith an additional suitable therapeutic agent, for example, acancer-therapeutic agent. When combination therapy is employed, aneffective amount can be achieved using a first amount of a compound ofpresent invention or a pharmaceutically acceptable salt thereof and asecond amount of an additional suitable therapeutic agent.

In one embodiment, the compound of present invention and the additionaltherapeutic agent, are each administered in an effective amount (i.e.,each in an amount which would be therapeutically effective ifadministered alone). In another embodiment, the compound of presentinvention and the additional therapeutic agent, are each administered inan amount which alone does not provide a therapeutic effect (asub-therapeutic dose). In yet another embodiment, the compound ofpresent invention can be administered in an effective amount, while theadditional therapeutic agent is administered in a sub-therapeutic dose.In still another embodiment, the compound of present invention can beadministered in a sub-therapeutic dose, while the additional therapeuticagent, for example, a suitable cancer-therapeutic agent is administeredin an effective amount.

As used herein, the terms “in combination” or “coadministration” can beused interchangeably to refer to the use of more than one therapies(e.g., one or more prophylactic and/or therapeutic agents). The use ofthe terms does not restrict the order in which therapies (e.g.,prophylactic and/or therapeutic agents) are administered to a subject.

Coadministration encompasses administration of the first and secondamounts of the compounds of the coadministration in an essentiallysimultaneous manner, such as in a single pharmaceutical composition, forexample, capsule or tablet having a fixed ratio of first and secondamounts, or in multiple, separate capsules or tablets for each. Inaddition, such coadministration also encompasses use of each compound ina sequential manner in either order.

When coadministration involves the separate administration of the firstamount of a compound of present invention and a second amount of anadditional therapeutic agent, the compounds are administeredsufficiently close in time to have the desired therapeutic effect. Forexample, the period of time between each administration which can resultin the desired therapeutic effect, can range from minutes to hours andcan be determined taking into account the properties of each compoundsuch as potency, solubility, bioavailability, plasma half-life andkinetic profile. For example, a compound of present invention and thesecond therapeutic agent can be administered in any order within about24 hours of each other, within about 16 hours of each other, withinabout 8 hours of each other, within about 4 hours of each other, withinabout 1 hour of each other or within about 30 minutes of each other.

More, specifically, a first therapy (e.g., a prophylactic or therapeuticagent such as a compound of the invention) can be administered prior to(e.g., 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4hours, 6 hours, 12 hours, 24 hours, 48 hours, 72 hours, 96 hours, 1week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks, or 12 weeksbefore), concomitantly with, or subsequent to (e.g., 5 minutes, 15minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12hours, 24 hours, 48 hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks,4 weeks, 5 weeks, 6 weeks, 8 weeks, or 12 weeks after) theadministration of a second therapy (e.g., a prophylactic or therapeuticagent such as an anti-cancer agent) to a subject.

It is understood that the method of coadministration of a first amountof a compound of present invention and a second amount of an additionaltherapeutic agent can result in an enhanced or synergistic therapeuticeffect, wherein the combined effect is greater than the additive effectthat would result from separate administration of the first amount ofthe compound of present invention and the second amount of theadditional therapeutic agent.

As used herein, the term “synergistic” refers to a combination of acompound of the invention and another therapy (e.g., a prophylactic ortherapeutic agent), which is more effective than the additive effects ofthe therapies. A synergistic effect of a combination of therapies (e.g.,a combination of prophylactic or therapeutic agents) permits the use oflower dosages of one or more of the therapies and/or less frequentadministration of said therapies to a subject. The ability to utilizelower dosages of a therapy (e.g., a prophylactic or therapeutic agent)and/or to administer said therapy less frequently reduces the toxicityassociated with the administration of said therapy to a subject withoutreducing the efficacy of said therapy in the prevention, management ortreatment of a disorder. In addition, a synergistic effect can result inimproved efficacy of agents in the prevention, management or treatmentof a disorder. Finally, a synergistic effect of a combination oftherapies (e.g., a combination of prophylactic or therapeutic agents)may avoid or reduce adverse or unwanted side effects associated with theuse of either therapy alone.

The presence of a synergistic effect can be determined using suitablemethods for assessing drug interaction. Suitable methods include, forexample, the Sigmoid-Emax equation (Holford, N. H. G. and Scheiner, L.B., Clin. Pharmacokinet. 6: 429-453 (1981)), the equation of Loeweadditivity (Loewe, S. and Muischnek, H., Arch. Exp. Pathol Pharmacol.114: 313-326 (1926)) and the median-effect equation (Chou, T. C. andTalalay, P., Adv. Enzyme Regul. 22: 27-55 (1984)). Each equationreferred to above can be applied with experimental data to generate acorresponding graph to aid in assessing the effects of the drugcombination. The corresponding graphs associated with the equationsreferred to above are the concentration-effect curve, isobologram curveand combination index curve, respectively.

The activity of the compounds as inhibitors of bacteria infection may beassayed in vitro or in vivo. In vitro assays include assays thatdetermine inhibition of FimH activity, bacterial adhesion, and bacterialbinding. Alternate in vitro assays quantitate the ability of theinhibitor to bind to the FimH and may be measured either byradiolabelling the inhibitor prior to binding, isolating the inhibitorcomplex and determining the amount of radiolabel bound, or by running acompetition experiment where new inhibitors are incubated with the FimHbound to known radioligands. Detailed conditions for assaying a compoundutilized in this invention are set forth in the Examples below.

Experimental Details

The following abbreviations are used in the examples below:

-   Ac acetyl-   AcOH acetic acid-   AC₂O acetic anhydride-   BF₃.OEt₂ diethyloxonio-trifluoro-boron-   Bn benzyl-   CH₃CN acetonitrile-   CD₃OD methanol-D4-   CDCl₃ chloroform-D-   CH₂C₁₂ methylene chloride or dichloromethane-   conc concentrate-   Cs₂CO₃ cesium carbonate-   Cut copper(I) iodide-   CuSO₄ copper(II) sulfate-   CV column volume-   DBU 1,8-diazabicyclo[5.4.0]undec-7-ene-   DIPEA N-ethyl-N-isopropyl-propan-2-amine-   DMAP 4-dimethylaminopyridine-   DMF dimethylformamide-   DMSO dimethylsulfoxide-   Eq. equivalent-   EtOAc ethyl acetate-   HATU    O-(7-azabenzotriazol-1-yl),N,N,N″,N″-tetramethyluroniumhexafluorophosphate-   h hour(s)-   Hex hexanes-   M molar-   MeOH methanol-   MeONa sodium methoxide-   min minute(s)-   MTBE methyl tert-butyl ether-   NaIO₄ sodium periodate-   Na₂SO₄ sodium sulfate-   NMO N-methylmorpholine-N-oxide-   OsO₄ osmium tetroxide-   PdCl₂ palladium (II)chloride-   Pd(PPh₃)₄ palladium tetrakis triphenylphosphine-   Pd(OAc)₂ palladium(II)acetate-   PdCl₂(dppf).CH₂Cl₂    (1,1′-Bis-(diphenylphosphino)-ferrocene)palladium (II) dichloride    dichloromethane complex-   Pd(OH)₂ dihydroxy palladium-   Piv trimethylacetyl-   Py pyridine-   RBF round bottom flask-   RT room temperature-   TBAF tetrabutylammonium fluoride-   TBDMSOTf tert-butyldimethylsilyl trifluoromethanesulfonate-   TBS tert-butyldimethylsilyl-   TEA triethylamine-   Tf trifluoromethanesulfonyl-   TFA trifluoroacetic acid-   THF tetrahydrofuran-   TLC thin layer chromatography-   TMS trimethylsilyl-   TMSI trimethylsilyl iodide-   TMSN₃ trimethylsilyl azide-   TMSOTf trimethylsilyl trifluoromethanesulfonate

The compounds of this invention may be prepared in light of thespecification using steps generally known to those of ordinary skill inthe art. Those compounds may be analyzed by known methods, including butnot limited to LC-MS (liquid chromatography mass spectrometry), HPLC(high performance liquid chromatography) and NMR (nuclear magneticresonance). It should be understood that the specific conditions shownbelow are only examples, and are not meant to limit the scope of theconditions that can be used for making compounds of this invention.Instead, this invention also includes conditions that would be apparentto those skilled in that art in light of this specification for makingthe compounds of this invention. Unless otherwise indicated, allvariables in the following Examples are as defined herein.

Mass spec. samples are analyzed on a Waters UPLC Acquity massspectrometer operated in single MS mode with electrospray ionization.Samples are introduced into the mass spectrometer using chromatography.Mobile phase for the mass spec. analyses consisted of 0.1% formic acidand CH₃CN-water mixture. Column gradient conditions are 5%-85%CH₃CN-water over 6 minutes run time, AcquityHSS T3 1.8μ, 2.1 mm ID×50 mmFlow rate is 1.0 mL/min. As used herein, the term “Rt(min)” refers tothe LC-MS retention time, in minutes, associated with the compound.Unless otherwise indicated, the LC-MS method utilized to obtain thereported retention time is as detailed above.

Purification by reverse phase HPLC is carried out under standardconditions using either Phenomenex Gemini 21.2 mm ID×250 mm column (5μm), Gemini 21.2 mm ID×75 mm column, (5 μm), 110 Å or in most cases aWaters XSELECT CSH Prep C18 (5 μm) ODB 19×100 mm column. Elution isperformed using a linear gradient CH₃CN—H₂O (with or without 0.01% TFAbuffer or 0.1% HCOH) as mobile phase. Solvent system is tailoredaccording to the polarity of the compound, Flow rate, 20 mL/minCompounds are collected either by UV or Waters 3100 Mass Detector, ESIPositive Mode. Fractions containing the desired compound are combined,concentrated (rotary evaporator) to remove excess CH₃CN and theresulting aqueous solution is lyophilized to afford the desired materialin most cases as a white foam.

General Method of Synthesis

Compounds described therein are prepared from key intermediates usingtwo key reactions; Suzuki and Sonogashira coupling.

Compounds of formula IV (Z=i) can be prepared by two distinct methods,as exemplified in Scheme 1. In Method A, a palladium catalyzedstereoselective C-glycosidation of per-acetylated glycal witharyl/heteroaryl bis-boronic of type I generates selectively bis-α-glucalof type II (Maddaford et al. Org Letters, 2001, 3 (13), 2013). Astereoselective bis-dihydroxylation of the latter (OsO₄, NMO) affordsthe aryl/heteroaryl bis-α-mannosides of type III. Removal of the acetateprotective group by saponification (MeONa/MeOH) generates the desiredfinal product of type IV (Z=i). Alternatively in Method B, thestereoselective C-glycosidation results from the double addition ofaryl/heteroaryl zinc reagents derived from diiodo aryl/heteroaryl oftype V on(3S,4S,5R,6R)-2-bromo-6-((pivaloyloxy)methyl)tetrahydro-2H-pyran-3,4,5-triyltris(2,2-dimethylpropanoate) as reported by Knochel (Org Letters, 2012,14 (6), 1480). The resulting fully pivalated bismannosides VI isdeprotected under acidic condition (AcOH, THF, H₂O) to afford thedesired material IV.

Compounds of formula XI (Z=ii), XIV (Z=iii), and XIX (Z=vii) can also beprepared using Methods A and B, as exemplified in Scheme 2. Biaryls orbis-heteroaryls of type XI can either be prepared sequentially in MethodA for primarily unsymmetrical analogs (B≠C) or more promptly usingMethod B, pending the availability of starting material of type V. InMethod A, intermediates of type IX are prepared as described previouslyusing boronic acids of type VII. In some cases, the boronic acid VII isreplaced by the corresponding pinacolo-boronate prepared from thecorresponding benzyl protected bromo or iodo-phenol. After coupling withthe glucal and dihydroxylation the benzyl protective group is removed byhydrogenolysis and converted to the triflate to enable palladiumcatalyzed cross coupling. Suzuki coupling of intermediates IX and Xfollowed by saponification provides the desired bis-mannosides XI.Compounds of type XIV (Z=iii) can be prepared in a similar fashion fromthe pinacol boronate of type XII by Suzuki coupling with bis halogenatedintermediates of type XIII This approach is performed in two steps(coupling and deprotection) for symmetrical compounds (D=F) orsequentially for unsymmetrical (D≠F) analogs. In Method B, compounds oftypes XI, XIV and XIX are prepared as described previously fromintermediates XV, XVI and XVIII respectively either commerciallyavailable or prepared by cross coupling strategies.

Compounds of formula XXII (Z=v) in which L2 and L3 are alkyne, can beprepared by three distinct Methods (C, D and E), as exemplified inScheme 3. In Method C, the intermediate XXI is prepared via aSonogashira coupling between dibromo or diiodo aryl or heteroaryl oftype XX and((2R,3R,4R,5R,6R)-6-ethynyl-3,4,5-tris((triisopropylsilyl)oxy)tetrahydro-2H-pyran-2-yl)methanolor the TMS protected analog (see Helv. Chim. Acta. 2001, 84(8),2355-2367). Removal of the TIPS protecting groups of XVIII under acidiccondition (TFA, THF, H₂O) or using TBAF affords the desired compoundXXII. Alternatively, in Method D, the same Sonogashira couplingdescribed in Method C is performed using the unprotected α-ethynylmannose((2R,3S,4R,5S,6R)-2-ethynyl-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol;Helv. Chim. Acta. 2001, 84(8), 2355-2367). Finally in Method E, a Lewisacid catalyzed double Ferrier type alkynylation between(2R,3S,4R)-2-(acetoxymethyl)-3,4-dihydro-2H-pyran-3,4-diyl diacetate andthe bis-TMS-acetylene aryl of heteroaryl type XXIII compound can beperformed to afford the desired intermediate XXIV (Tetrahedron Letters2002, 43, 5437-5440. A stereospecific bis-dihydroxylation ofintermediate XXIV followed by a saponification generated the desiredfinal compound XXII.

Bis-mannosides of type XXVII exhibiting α-aryl and α-alkynyl linkers tomannose are prepared by reactions previously described, according to thesynthetic route of Scheme 4, Method F. Sonogashira coupling betweenhalogenated aryls or heteroaryls of type XXVI (prepared as described forIX) and(2R,3R,4R,5R,6R)-6-ethynyl-3,4,5-tris((triisopropylsilyl)oxy)tetrahydro-2H-pyran-2-yl)methanolor the TMS protected analog can afford after sequential deprotection ofacetate and TIPS groups if necessary the desired compounds of typeXXVII.

The following is a list of key intermediates which are used in thepreparation of Compounds described therein

Preparation of Intermediate A

([(2R,3S,4R,5S,6R)-3-Acetoxy-4,5-dihydroxy-6-[4-(trifluoromethylsulfonyloxy)phenyl]tetrahydropyran-2-yl]methylacetate)

Step I:[(2R,3S,6S)-3-Acetoxy-6-(4-hydroxyphenyl)-3,6-dihydro-2H-pyran-2-yl]methylAcetate

Acetonitrile (50.00 mL) is added to a mixture of[(2R,3S,4R)-3,4-diacetoxy-3,4-dihydro-2H-pyran-2-yl]methyl acetate(9.869 g, 36.25 mmol), (4-hydroxyphenyl)boronic acid (5 g, 36.25 mmol)and Pd(OAc)₂ (1.221 g, 5.438 mmol) and the reaction mixture is stirredat RT overnight. An additional amount of (4-hydroxyphenyl)boronic acid(1 g) is added and the reaction mixture is stirred for a further 2 h andfiltered through celite. The filtrate is evaporated and the crudeproduct is purified on a Biotage™ SNAP 340 g silica gel cartridge with agradient of 5%-80% EtOAc in Hex to afford title product (6.03 g).

Step II:[(2R,3S,4R,5S,6R)-3-Acetoxy-4,5-dihydroxy-6-(4-hydroxyphenyl)tetrahydropyran-2-yl]methylAcetate

To a suspension of[(2R,3S,6S)-3-acetoxy-6-(4-hydroxyphenyl)-3,6-dihydro-2H-pyran-2-yl]methylacetate (6.03 g, 19.69 mmol) in THF (36 mL)/water (24 mL) are addedmethanesulfonamide (2.810 g, 29.54 mmol), OsO₄ (6.007 g, 7.4 mL of 2.5%w/w in -t-BuOH, 0.5907 mmol) and NMO (4.613 g, 39.38 mmol). The reactionmixture is stirred at RT overnight. 1M Na₂S₂O₃ (40 mL) is added and themixture is extract with EtOAc (3×40 mL). The combined organic extractsare washed with brine (15 mL) and dried over Na₂SO₄. The mixture isfiltered, the solvent is evaporated and the crude product is purified ona Biotage™ SNAP 220 g silica gel cartridge with a gradient of 0%-20%MeOH in CH₂Cl₂ to afford title product (6.05 g).

LC-MS: m/z=329.3 (M+Na⁺)

Step III: Intermediate A

To a solution of[(2R,3S,4R,5S,6R)-3-acetoxy-4,5-dihydroxy-6-(4-hydroxyphenyl)tetrahydropyran-2-yl]methylacetate (872 mg, 2.562 mmol) in CH₂Cl₂ (22 mL) are added1,1,1-trifluoro-N-phenyl-N-(trifluoromethylsulfonyl)methanesulfonamide(1.190 g, 3.331 mmol), NEt₃ (518.5 mg, 714 μL, 5.124 mmol) and thereaction mixture is stirred at RT overnight. The solvent is evaporatedand the crude product is purified on a Biotage™ SNAP 100 g silica gelcartridge with a gradient of 0%-20% MeOH/CH₂Cl₂ over 15 column volume toafford the title product (1.06 g).

Preparation of Intermediate B

([(2R,3S,4R,5S,6R)-3-Acetoxy-4,5-dihydroxy-6-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]tetrahydropyran-2-yl]methylacetate)

A mixture of[(2R,3S,4R,5S,6R)-3-acetoxy-4,5-dihydroxy-6-[4-(trifluoromethylsulfonyloxy)phenyl]tetrahydropyran-2-yl]methylacetate (350 mg, 0.74 mmol),4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolane(225.8 mg, 0.89 mmol), PdCl₂(dppf). CH₂Cl₂ (45.18 mg, 0.074 mmol) andpotassium acetate (291 mg, 2.96 mmol) in DMF (3.7 mL) is degassed bybubbling N₂(g) through for 5 min. The reaction mixture is then heated at80° C. for 5h. The solvent is removed under high vacuum and the crudeproduct is triturated with EtOAc (10 mL) and filtered. The filtrate isconcentrated and purified on a Biotage™ SNAP 25 g silica gel cartridgewith a gradient elution of 40%-60% EtOAc in Hex and a flow rate of 24mL/min over 20 min to afford the title product (168 mg, 0.3731 mmol,50.36%) as a foam. ¹H NMR (400 MHz, CD₃OD) δ 7.75 (d, J=7.8 Hz, 2H),7.45 (d, J=7.9 Hz, 2H), 4.96-4.90 (m, 1H), 4.72 (dd, J=12.1, 8.2 Hz,1H), 4.20-4.02 (m, 2H), 3.93-3.76 (m, 2H), 3.49 (dd, J=14.4, 7.0 Hz,1H), 2.07 (s, 3H), 2.05 (s, 3H), 1.35 (s, 12H). LC-MS: m/z=451.3 (M+H⁺).

Preparation of Intermediate C

((2R,3S,4R,5S,6R)-2-(3-Bromophenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol)

Step I:((2R,3S,6S)-3-Acetoxy-6-(3-bromophenyl)-3,6-dihydro-2H-pyran-2-yl)methylacetate

A solution of [(2R,3S,4R)-3,4-diacetoxy-3,4-dihydro-2H-pyran-2-yl]methylacetate (3.00 g, 11.02 mmol) and (3-bromophenyl)boronic acid (4.43 g,22.04 mmol) in CH₃CN (22 mL) is degassed by bubbling nitrogen gasthrough for 3 min. Palladium (II) acetate (371 mg, 1.65 mmol) is addedand the reaction mixture is stirred at RT for 5h then another portion ofpalladium (II) acetate (371 mg, 1.65 mmol) is added and stirring iscontinued for 18 h. The solvent is evaporated and the mixture is dilutedwith dichloromethane (10 mL) and saturated aqueous NaHCO₃ (20 mL). Themixture is filtered through a phase separator cartridge, the filtrate isevaporated and purified on a Biotage™ SNAP 50 g silica gel cartridgeusing a gradient elution of 5%-10% EtOAc/Hex with a flow rate of 40mL/min over 30 min to afford title product (1.61 g, 4.36 mmol, 40%) asan oil. LC-MS: m/z=391.1, 393.1 (M+Na⁺)

Step II:((2R,3S,4R,5S,6R)-3-Acetoxy-6-(3-bromophenyl)-4,5-dihydroxytetrahydro-2H-pyran-2-yl)methylAcetate

To a solution of[(2R,3S,6S)-3-acetoxy-6-(3-bromophenyl)-3,6-dihydro-2H-pyran-2-yl]methylacetate (1.60 g, 4.33 mmol) in water (3 mL) and THF (19 mL) is addedmethanesulfonamide (618 mg, 6.50 mmol), osmium tetroxide (1.3 mL of 2.5%w/v in t-BuOH, 0.130 mmol) and N-methylmorpholine-N-oxide (2.030 g,17.33 mmol) and the reaction mixture is stirred at RT for 2 days.Another portion of osmium tetroxide (1.3 mL of 2.5% w/v in t-BuOH, 0.130mmol), methanesulfonamide (618 mg, 6.50 mmol) andN-methylmorpholine-N-oxide (2.030 g, 17.33 mmol) are added and themixture is stirred for a further 24 h. The solvent is evaporated and thecrude mixture is diluted with a saturated solution of sodium bisulfite(50 mL) and extracted with EtOAc (3×15 mL). The combined organicextracts are dried over Na₂SO₄ and the solvent is evaporated. Thegel-like material obtained is dissolved in a minimum amount of MeOH anddiluted with diethyl ether and placed in the fridge for 2 h. The mixtureis filtered and washed with diethyl ether and dried under high vacuum toafford title product (1.480 g, 85%) as a solid. LC-MS: m/z=425.1, 427.1(M+Na⁺)

Step III: Intermediate C

((2R,3S,4R,5S,6R)-3-Acetoxy-6-(3-bromophenyl)-4,5-dihydroxytetrahydro-2H-pyran-2-yl)methylacetate (1.48 g) is dissolved in MeOH (20 mL) and MeONa in MeOH (187 μLof 25% w/v, 0.87 mmol) is added and the reaction mixture is stirred atRT for 4h. The reaction mixture is neutralized by the addition ofAmberlite IR120H resin until the pH changed to neutral. The reactionmixture is filtered, the filtrate is evaporated and the solid istriturated with Et₂O (2×10 mL) to afford the title product (1.08 g,3.046 mmol, 70.3%) as a solid. ¹H NMR (400 MHz, CD₃OD) δ 7.68 (s, 1H),7.45 (dd, J=10.6, 4.1 Hz, 2H), 7.29 (t, J=7.9 Hz, 1H), 4.91 (d, J=4.7Hz, 1H), 4.29 (dd, J=4.6, 3.2 Hz, 1H), 3.95-3.71 (m, 3H), 3.61 (dd,J=7.4, 3.1 Hz, 1H), 3.55-3.47 (m, 1H). LC-MS: m/z=341.1, 343.1 (M+Na⁺).

Preparation of Intermediate D

3-((2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)benzoicAcid

Step I: methyl3-((2S,5S,6R)-5-acetoxy-6-(acetoxymethyl)-5,6-dihydro-2H-pyran-2-yl)benzoate

Methyl3-[(2R,3S,6S)-3-acetoxy-2-(acetoxymethyl)-3,6-dihydro-2H-pyran-6-yl]benzoateis prepared using the same procedure as described in Step I for thepreparation of Intermediate C, but using(3-methoxycarbonylphenyl)boronic acid as the starting material.

LC-MS: m/z=371.2 (M+Na⁺).

Step II: methyl3-((2R,3S,4R,5S,6R)-5-acetoxy-6-(acetoxymethyl)-3,4-dihydroxytetrahydro-2H-pyran-2-yl)benzoate

Methyl3-[(2R,3S,4R,5S,6R)-5-acetoxy-6-(acetoxymethyl)-3,4-dihydroxy-tetrahydropyran-2-yl]benzoateis prepared using the same procedure as described in Step II for thepreparation of Intermediate C. LC-MS: m/z=383.3 (M+H⁺).

Step III: Intermediate D

A mixture of methyl3-[(2R,3S,4R,5S,6R)-5-acetoxy-6-(acetoxymethyl)-3,4-dihydroxy-tetrahydropyran-2-yl]benzoate(2.20 g, 5.75 mmol) in MeOH (30 mL) is treated with MeONa in MeOH (341μL of 25% w/v, 1.58 mmol) and the reaction mixture is stirred at RT for18h. The volatiles are evaporated, the mixture is dissolved in MeOH (30mL), aqueous sodium hydroxide (5.1 mL of 2 M, 10.3 mmol) is added andthe reaction mixture is stirred at RT for 15h. The mixture isneutralized by the addition of Amberlite IR120H resin until the pHchanged to neutral. The reaction mixture is filtered and the filtrate isevaporated to afford the title product (1.57 g, 66%) as a white solid.¹H NMR (400 MHz, CD₃OD) δ 8.15 (s, 1H), 7.96 (d, J=7.7 Hz, 1H), 7.75 (d,J=7.7 Hz, 1H), 7.51 (t, J=7.8 Hz, 1H), 4.99 (d, J=4.4 Hz, 1H), 4.42-4.36(m, 1H), 3.93-3.74 (m, 3H), 3.63 (dd, J=7.5, 3.1 Hz, 1H), 3.54-3.49 (m,1H). LC-MS: m/z=285.2 (M+H⁺).

Preparation of Intermediate E

((2R,3R,4R,5R,6R)-2-(Acetoxymethyl)-6-(3-hydroxyphenyl)tetrahydro-2H-pyran-3,4,5-triyltriacetate)

Step I:[(2R,3S,6S)-3-Acetoxy-6-[3-[tert-butyl(dimethyl)silyl]oxyphenyl]-3,6-dihydro-2H-pyran-2-yl]methylAcetate

To a solution of[(2R,3S,4R)-3,4-diacetoxy-3,4-dihydro-2H-pyran-2-yl]methyl acetate(1.100 g, 4.040 mmol) in 10 mL of CH₃CN are added[3-(tert-butyl-dimethyl-silyl)oxyphenyl]boronic acid (2.038 g, 8.080mmol) and Pd(OAc)₂ (136.1 mg, 0.6060 mmol). The mixture is stirred at RTfor 5 h and then to it are added another batch of Pd(OAc)₂ (136 mg,0.606 mmol) and [3-(tert-butyl-dimethyl-silyl)oxyphenyl]boronic acid(2.038 g, 8.080 mmol). It is then stirred at RT overnight. The mixtureis diluted with 20 mL of CH₂Cl₂ and filtered over a pad of celite. Thefiltrate is concentrated and the residue is separated on Biotage™ SNAP100 g silica gel cartridge using a gradient of EtOAc in Hex (0-20%) in20 column volume to afford the title product (805 mg, 47%) as an oil,which solidifies upon standing. ¹H NMR (CDCl₃, 400 MHz): 7.06 (m, 1H),6.78 (m, 1H), 6.70 (m, 1H), 6.60 (m, 1H), 5.97 (m, 1H), 5.71 (m, 1H),5.09 (m, 2H), 4.08 (m, 1H), 3.85 (m, 1H), 3.62 (m, 1H), 1.88 and 1.87(2s, 6H), 0.78 (m, 9H), 0.00 (m, 6H).

Step II:[(2R,3S,4R,5S,6R)-3-Acetoxy-6-[3-[tert-butyl(dimethyl)silyl]oxyphenyl]-4,5-dihydroxy-tetrahydropyran-2-yl]methylAcetate

To a solution of[(2R,3S,6S)-3-acetoxy-6-[3-[tert-butyl(dimethyl)silyl]oxyphenyl]-3,6-dihydro-2H-pyran-2-yl]methylacetate (2.500 g, 5.944 mmol) in water (10 mL)/t-BuOH (10 mL) are addedmethanesulfonamide (848.0 mg, 8.92 mmol), 2.5% OsO₄/t-BuOH (1.9 mL,0.149 mmol), NMO (1.393 g, 11.89 mmol) and lutidine (689 μL, 5.94 mmol).The mixture is stirred at RT overnight. It is then quenched with 15%sodium bisulfate (15 mL) and diluted with EtOAc (40 mL). The aqueousphase is then separated, washed with water (20 mL) and brine (20 mL)consecutively, dried over Na₂SO₄. After removal of the solvent underreduced pressure, the residue is purified on Biotage™ SNAP 100 g silicagel cartridge using a gradient of MeOH in CH₂Cl₂ (0-6%) in 20 columnvolume to afford the title compound (2.20 g, 81%) as an oil. ¹H NMR(CD₃OD, 400 MHz): 7.06 (m, 1H), 6.78 (m, 1H), 6.70 (m, 1H), 6.58 (m,1H), 4.85 (m, 1H), 4.64 (m, 1H), 4.46 (m, 1H), 3.96 (m, 1H), 3.85 (m,1H), 3.62 (m, 2H), 1.86 and 1.83 (2s, 6H), 0.78 (m, 9H), 0.00 (m, 6H).

Step III:(2R,3R,4R,5R,6R)-2-(Acetoxymethyl)-6-(3-((tert-butyldimethylsilyl)oxy)phenyl)tetrahydro-2H-pyran-3,4,5-triyltriacetate

To a solution of[(2R,3S,4R,5S,6R)-3-acetoxy-6-[3-[tert-butyl(dimethyl)silyl]oxyphenyl]-4,5-dihydroxy-tetrahydropyran-2-yl]methylacetate (1.00 g, 2.20 mmol) in methylene chloride (20 mL) at 0° C.,under N₂, is added 2,6-lutidine (872 μL, 6.60 mmol) followed by DMAP(53.8 mg, 0.44 mmol) & acetic anhydride (623 μL, 6.60 mmol). The yellowsolution is stirred at 0° C. for 1.5 h. TLC (30% EtOAc/hex) showedcomplete consumption of starting material. The reaction is treated withKHSO₄ (15%, 2×6 mL) then washed with brine, dried & evaporated. Thecrude material is purified on Biotage™ SNAP silica gel cartridge usingEtOAc-Hex (0-5%, 3 CV; 5-30%, 20 CV) as eluent to afford the titleproduct (1.02 g) as a clear gum.

Step IV: Intermediate E

To a stirred solution of[(2R,3R,4R,5R,6R)-3,4,5-triacetoxy-6-[3-[tert-butyl(dimethyl)silyl]oxyphenyl]tetrahydropyran-2-yl]methylacetate (1.02 g, 1.799 mmol) in THF (10 mL) are added acetic acid (162.0mg, 153 μL, 2.698 mmol) and TBAF (5.4 mL of 1 M, 5.397 mmol). Themixture is stirred at RT for 30 min. It is then diluted with EtOAc(30mL), washed with water (20 mL) and brine (20 mL), dried over sodiumsulfate, and concentrated. The crude material is purified on Biotage™SNAP 25 g silica gel column using EtOAc in Hex (5%, 5 CV; 5-30%, 25 CV;30-40% 5 CV, 40-50%, 30 CV) as eluent to afford the title product (409mg, 48.35%) as white foam. ¹H NMR (400 MHz, CD₃OD) δ 7.23 (t, 1H), 6.92(dd, 2H), 6.76 (dd, 1H), 5.86 (t, 1H), 5.24 (t, 1H), 5.10 (dd, 1H), 5.01(d, 1H), 4.37 (dd, 1H), 4.13 (dd, 1H), 3.86-3.66 (m, 1H), 2.08 (d, 6H),2.02 (d, 6H).

Preparation of Intermediate F

((2R,3R,4R,5R,6R)-2-(Acetoxymethyl)-6-(4-ethynylphenyl)tetrahydro-2H-pyran-3,4,5-triyltriacetate)

Step I:[(2R,3R,4R,5R,6R)-3,4,5-Triacetoxy-6-[4-(trifluoromethylsulfonyloxy)phenyl]tetrahydropyran-2-yl]methylAcetate

To a solution of(2R,3S,4R,5S,6R)-3-acetoxy-4,5-dihydroxy-6-[4-(trifluoromethylsulfonyloxy)phenyl]tetrahydropyran-2-yl]methylacetate (256 mg, 0.542 mmol) in 2.6 mL of CH₂Cl₂ is sequentially addedpyridine (132 μL, 1.63 mmol), Ac₂O (128 μL, 1.36 mmol) and DMAP (6.6 mg,0.054 mmol). The reaction mixture is stirred at RT for 2 h, diluted withwater (1 mL) and the organic layer is dried over Na₂SO₄, filtered, andconcentrated to dryness. The residue is purified by flash columnchromatography on silica gel (10 to 80% EtOAc in Hex) to give titleproduct (232 mg, 77%).

Step II:(2R,3R,4R,5R,6R)-2-(Acetoxymethyl)-6-(4-((trimethylsilyl)ethynyl)phenyl)tetrahydro-2H-pyran-3,4,5-triylTriacetate

To a mixture of[(2R,3R,4R,5R,6R)-3,4,5-triacetoxy-6-[4-(trifluoromethylsulfonyloxy)phenyl]tetrahydropyran-2-yl]methylacetate (1217 mg, 2.187 mmol), Pd(dppf)Cl₂— CH₂Cl₂ (178.6 mg, 0.2187mmol) and CuI (83.3 mg, 0.437 mmol) in 12 mL of DMF is added Et₃N (1.5mL, 11 mmol) followed by ethynyl(trimethyl)silane (1.54 mL, 10.9 mmol).The reaction mixture is heated at 70° C. in a sealed tube for 21 h,cooled to RT, and diluted with water (40 mL). The reaction mixture isextracted by EtOAc (5×20 mL), and the combined organic layer are washedwith water (3×10 mL), 10 mL of brine, dried over Na₂SO₄, filtered, andconcentrated to dryness. The residue is purified by flash columnchromatography on silica gel (10 to 80% EtOAc in Hex) to give titleproduct (1.0596 g, 96%).

Step III: Intermediate F

To a solution of(2R,3R,4R,5R,6R)-2-(acetoxymethyl)-6-(4-((trimethylsilyl)ethynyl)phenyl)tetrahydro-2H-pyran-3,4,5-triyltriacetate (1.054 g, 2.089 mmol) in 21 mL of THF is sequentially addedAcOH (150.6 mg, 143 μL, 2.507 mmol) and TBAF 1M in THF (2.298 mL of 1 M,2.298 mmol) under nitrogen atmosphere. The reaction mixture is stirredat RT for 2 h, and concentrated to dryness. The residue is purified byflash column chromatography on silica gel (10 to 80% EtOAc in Hex) togive the title compound (892 mg, 99%).

Preparation of Intermediate G

((2R,3R,4R,5R,6R)-3,4,5-Tribenzyloxy-2-(benzyloxymethyl)-6-ethynyl-tetrahydropyran)

Step I:(2R,3R,4S,5S,6R)-3,4,5-tribenzyloxy-2-(benzyloxymethyl)-6-fluoro-tetrahydropyran

To a solution of(3S,4S,5R,6R)-3,4,5-tribenzyloxy-6-(benzyloxymethyl)tetrahydropyran-2-ol(10.8 g, 19.98 mmol) and (diethylamino)difluorosulfoniumtetrafluoroborate (7.075 g, 29.97 mmol) in 50 mL of CH₂Cl₂ is added DBU(4.8 mL, 32.1 mmol) at −15° C. and then stirred for 20 min. The reactionis quenched with saturated sodium bicarbonate solution. Then the mixtureis extracted with CH₂Cl₂ (3×20 mL). The combined organic extracts arewashed with water and brine consecutively, dried over sodium sulfate,filtered, and concentrated to dryness. The residue is separated onBiotage™ SNAP 100 g silica gel cartridge using a gradient of EtOAc inHex (0-15%, 20 CV) to obtain a major fraction containing title product(6.40 g). LC-MS: m/z=565.4 (M+Na⁺).

Step II:Trimethyl-[2-[(2R,3R,4R,5R,6R)-3,4,5-tribenzyloxy-6-(benzyloxymethyl)tetrahydropyran-2-yl]ethynyl]silane

To a solution of(2R,3R,4S,5S,6R)-3,4,5-tribenzyloxy-2-(benzyloxymethyl)-6-fluoro-tetrahydropyran(1120 mg, 2.064 mmol) and trifluoro(2-trimethylsilylethynyl)boranuide(Potassium Ion (1)) (547.6 mg, 2.683 mmol) in CH₃CN (15 mL) is addedBF₃.OEt₂ (351.6 mg, 306 μL, 2.48 mmol) at −10° C. and the mixture isstirred under nitrogen for 20 min at the same temperature. Then it isdiluted with EtOAc(30 mL), quenched with saturated sodium bicarbonatesolution, washed with water and brine consecutively, dried over sodiumsulfate, concentrated to dryness. The residue is separated on Biotage™SNAP 50 g silica gel cartridge using a gradient of EtOAc in Hex (0-15%,20 CV) to obtain title product (1.06 g) as oil.

Step III: Intermediate G

To a solution oftrimethyl-[2-[(2R,3R,4R,5R,6R)-3,4,5-tribenzyloxy-6-(benzyloxymethyl)tetrahydropyran-2-yl]ethynyl]silane(550 mg, 1.0 mmol) in THF (10 mL) is added 1M TBAF/THF (1.5 mL of 1 M,1.500 mmol). The mixture is stirred at RT for 20 min. Then it is dilutedwith EtOAc (30 mL), washed with water and brine consecutively, driedwith sodium sulfate, filtered and concentrated to dryness. The residueis separated on Biotage™ SNAP 25 g silica gel cartridge using a gradientof EtOAc in Hex 0-15% in 20 CV to obtain(2R,3R,4R,5R,6R)-3,4,5-tribenzyloxy-2-(benzyloxymethyl)-6-ethynyl-tetrahydropyran(450 mg) as an oil. ¹H NMR (400 MHz, CDCl₃) δ 7.44-7.20 (m, 18H),7.19-7.11 (m, 2H), 4.87 (d, 1H), 4.81 (t, 1H), 4.74 (d, 1H), 4.58 (m,6H), 4.09-3.91 (m, 3H), 3.85-3.68 (m, 3H), 2.49 (d, 1H).

Preparation of Intermediate H

([(2R,3R,4R,5R,6R)-3,4,5-Triacetoxy-6-allyl-tetrahydropyran-2-yl]methylacetate)

Step I:(2R,3R,4R,5R)-2-(Acetoxymethyl)-6-allyltetrahydro-2H-pyran-3,4,5-triylTriacetate

To a stirred solution of[(2R,3R,4S,5S,6R)-3,4,5,6-tetraacetoxytetrahydropyran-2-yl]methylacetate (5 g, 12.81 mmol) and allyl-trimethyl-silane (6.1 mL, 38.43mmol) in CH₃CN (30 mL) is added BF₃.EtO₂ (Ether (1)) (8.12 mL, 64.05mmol) at 0° C. The mixture is stirred at RT for 2 days, poured into asaturated solution of NaHCO₃ and stirred till bubbling stops. It is thenextracted with CH₂Cl₂. The combined organic extracts are washed withwater and brine consecutively, dried with sodium sulfate, filtered, andconcentrated to dryness. The residue is purified on Biotage™ SNAP 100 gsilica gel cartridge using a gradient of EtOAc in Hex 0-50% in 20 columnvolume to obtain title product (2.2 g, 46%), which contains α/β˜3:1. ¹HNMR (400 MHz, CDCl₃) δ 5.76 (ddt, 1H), 5.40-4.97 (m, 5H), 4.28 (ddd,1H), 4.21-3.96 (m, 2H), 3.96-3.81 (m, 1H), 2.57-2.33 (m, 2H), 2.18-1.94(m, 12H).

Step II:(3S,4R,5S,6R)-2-Allyl-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol

To a stirred solution of(2R,3R,4R,5R)-2-(acetoxymethyl)-6-allyltetrahydro-2H-pyran-3,4,5-triyltriacetate(3.9 g, 10.47 mmol) in MeOH (22.5 mL) is added 25% wt./v MeONa in MeOH(241 μL, 1.047 mmol). The mixture is stirred at RT overnight,neutralized with resin Amberlite 120 (H). After filtration, the filtrateis concentrated to dryness and the residue separated on Biotage™ SNAP 50g silica gel cartridge using a gradient of MeOH in CH₂Cl₂ (0-20%) in 24column volume to obtain title product (1.9 g, 89%). ¹H NMR (400 MHz,CD₃OD) δ 5.95-5.69 (m, 1H), 5.08 (ddd, 2H), 3.94-3.81 (m, 1H), 3.81-3.54(m, 5H), 3.50-3.37 (m, 1H), 2.53-2.40 (m, 1H), 2.34 (dq, 1H).

Step III:[(2R,3R,4R,5R,6R)-2-Allyl-3,5-bis[[tert-butyl(dimethyl)silyl]oxy]-6-[[tert-butyl(dimethyl)silyl]oxymethyl]tetrahydropyran-4-yl]oxy-tert-butyl-dimethyl-silane

To(3S,4R,5S,6R)-2-allyl-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol(1.9 g, 9.30 mmol) in CH₂Cl₂ (90 mL) is added DIPEA (8.10 mL, 46.52mmol) followed by TBDMSOTf (9.40 mL, 40.94 mmol). The reaction mixtureis stirred at RT overnight, then diluted with CH₂Cl₂, washed with asaturated solution of CuSO₄, H₂O, and brine. The organic phase is driedover Na₂SO₄, filtered and dried. The crude residue is purified byBiotage™ SNAP 100 g silica gel cartridge using EtOAc in Hex 0 to 2% in24CV to afford title product (4 g, 65%). ¹H NMR (400 MHz, CDCl₃) δ 5.89(tt, 1H), 5.18-4.83 (m, 2H), 3.98-3.82 (m, 2H), 3.82-3.67 (m, 4H), 3.63(t, 1H), 2.46 (dd, 1H), 2.00 (d, 1H), 1.10-0.66 (m, 36H), 0.28-−0.28 (m,24H).

Step IV: Intermediate H

To a solution of[(2R,3R,4R,5R,6R)-2-allyl-3,5-bis[[tert-butyl(dimethyl)silyl]oxy]-6-[[tert-butyl(dimethyl)silyl]oxymethyl]tetrahydropyran-4-yl]oxy-tert-butyl-dimethyl-silane(4 g, 6.05 mmol) in dry DMSO (35 mL) under N₂, is added TBAF (26.6 mL of1 M in THF, 26.62 mmol) and the RM is stirred for 3 h at 60° C. Thereaction is cooled down to RT, pyridine (5.38 mL, 66.54 mmol), aceticanhydride (5.7 mL, 60.49 mmol) and catalytic DMAP (36.94 mg, 0.30 mmol)are added and stirring is continued for 20h. The reaction is slowlypoured into ice/water and extracted with EtOAc. The organic layer iscarefully washed with a saturated solution of NaHCO₃, H₂O, and brine.The organic phase is dried over Na₂SO₄, filtered and dried to afford thetitle compound (2.00 g, 89%). ¹H NMR (400 MHz, CDCl₃) δ 5.76 (ddt, 1H),5.35-4.99 (m, 5H), 4.31 (dd, 1H), 4.20-3.94 (m, 2H), 3.94-3.71 (m, 1H),2.60-2.48 (m, 1H), 2.47-2.29 (m, 1H), 2.23-1.91 (m, 12H).

Preparation of Intermediate I

([(2R,3R,4R,5R,6R)-3,4,5-Triacetoxy-6-(3-bromophenyl)tetrahydropyran-2-yl]methylacetate)

To a solution of[(2R,3S,4R,5S,6R)-3-acetoxy-6-(3-bromophenyl)-4,5-dihydroxy-tetrahydropyran-2-yl]methylacetate (604.8 mg, 1.5 mmol) in THF (10 mL) are added DIPEA (969.3 mg,1.31 mL, 7.50 mmol), DMAP (18.3 mg, 0.150 mmol) and Ac₂O (536.0 mg, 495μL, 5.25 mmol) at 0° C. The mixture is stirred at RT overnight. Then itis quenched with saturated sodium bicarbonate solution. The mixture isextracted with CH₂Cl₂ (3×15 mL). The combined organic extracts arewashed with water and brine consecutively, dried over sodium sulfate,filtered, and concentrated to dryness. The residue is separated onBiotage™ SNAP 25 g silica gel cartridge using a gradient of EtOAc in Hex0-30% in 20 column volume to obtain title product (650 mg, 1.334 mmol,88.9%). ¹H NMR (400 MHz, CDCl₃) δ 7.66 (s, 1H), 7.55-7.45 (m, 1H), 7.41(dd, 1H), 7.29 (t, 1H), 5.87 (t, 1H), 5.28 (t, 1H), 5.10 (dd, 1H), 5.04(d, 1H), 4.36 (dd, 1H), 4.14 (dd, 1H), 3.86-3.66 (m, 1H), 2.13 (2s, 6H),2.05 (s, 3H), 2.02 (s, 3H)

Preparation of Intermediate J

(2-[(2R,3R,4R,5R,6R)-3,4,5-Triacetoxy-6-(acetoxymethyl)tetrahydropyran-2-yl]aceticacid)

To a solution of Intermediate H (220 mg, 0.590 mmol) in a mixture ofCH₃CN (1.1 mL)/CCl₄ (1.1 mL)/H₂O (1.9 mL) is added NaIO₄ (147.1 μL, 2.66mmol) followed by Cl₃Ru (Water (1)) (53.27 mg, 0.234 mmol). The reactionmixture is stirred at RT for 4 h, diluted with water and CH₂Cl₂,filtered on celite, washed with CH₂Cl₂. The aqueous phase is extractedtwice with CH₂Cl₂. Combined organic phases are dried over Na₂SO₄,filtered and concentrated to afford title product (110 mg, 74%) which isused in the next step without further purification. LC-MS: m/z=391.3(M+H⁺).

Preparation of Intermediate K

((2R,3S,4R,5S,6R)-2-(3-Ethynylphenyl)-6-(hydroxymethyl)tetrahydropyran-3,4,5-triol)

Step I:[(2R,3R,4R,5R,6R)-3,4,5-Tris(2,2-dimethylpropanoyloxy)-6-[3-(2-trimethylsilylethynyl)phenyl]tetrahydropyran-2-yl]methyl2,2-dimethylpropanoate

A solution of n-Bu₃MgLi (2.65 mL of 0.65 M, 1.725 mmol) inHex-heptane-dibutylether (8:20:3) is added to2-(3-bromophenyl)ethynyl-trimethyl-silane (1.248 g, 1.05 mL, 4.928 mmol)in toluene (2.4 mL) and dibutylether (1.4 mL) at 0° C. and stirred incold room for 25 h. A solution of ZnBr₂—LiBr in dibutyl ether (2.6 mL of1.05 M, 2.711 mmol) is added dropwise, cooling bath removed, stirred atRT for 1 h. A solution of[(2R,3R,4S,5S,6R)-6-bromo-3,4,5-tris(2,2-dimethylpropanoyloxy)tetrahydropyran-2-yl]methyl2,2-dimethylpropanoate (2.38 g, 4.107 mmol) in toluene (4.3 mL) isadded, it is placed on pre-heated oil bath at 90° C. for weekend. Thereaction mixture is cooled to RT, it is poured into aqueous 1 N HClsolution (40 mL) and extracted with EtOAc (3×40 mL). The combinedextracts are washed with brine, dried (Na₂SO₄), concentrated, purifiedon Biotage™ SNAP 100 g silica gel cartridge using EtOAc in Hex (0% to10%, 12 CV, 10%, 5 CV) as eluent to afford title product (765 mg) as anoil.

Step II: Intermediate K

To a stirred light suspension of[(2R,3R,4R,5R,6R)-3,4,5-tris(2,2-dimethylpropanoyloxy)-6-[3-(2-trimethylsilylethynyl)phenyl]tetrahydropyran-2-yl]methyl2,2-dimethylpropanoate (765 mg, 1.137 mmol) in MeOH (15 mL) is addedMeONa (4.6 mL of 0.5 M, 2.274 mmol) and stirred at RT for 24 h. To theresultant solution is added DOWEX 50WX4-400 until pH 4-5, filtered,eluted with MeOH. The filtrate is concentrated, purified on Biotage™SNAP 40 g silica gel cartridge using EtOAc-MeOH—H₂O (47.5:1.5:1 to10:1.5:1) as eluent to afford title product (170 mg, 55%) as beigesolid. LC-MS: m/z=265.28 (M+H⁺).

Preparation of Intermediate L

[(2R,3R,4R,5R,6R)-3,4,5-tris(triisopropylsilyloxy)-6-(2-trimethylsilylethynyl)tetrahydropyran-2-yl]methanol

Intermediate L is prepared according to the procedure described inJurgen Stichler-Bonaparte et. al. Helv. Chim. Acta. 2001, 84(8),2355-2367),

Preparation of Intermediate M

(2R,3S,4R,5S,6R)-2-ethynyl-6-(hydroxymethyl)tetrahydropyran-3,4,5-triol

Intermediate M is prepared according to the procedure described inJurgen Stichler-Bonaparte et. al. Helvetica Chimica Acta, 2001, 84(8),2355-2367

Preparation of Intermediate N

[(2R,3R,4R,5R,6R)-6-(4-bromophenyl)-3,4,5-tris(2,2-dimethylpropanoyloxy)tetrahydropyran-2-yl]methyl2,2-dimethylpropanoate

A solution of n-Bu₃MgLi (2.20 mL of 0.66 M, 1.45 mmol) inhexane-heptane-dibutyl ether (8:20:3) is added to 1-bromo-4-iodo-benzene(1.172 g, 4.141 mmol) in toluene (2.0 mL) and dibutyl ether (1.2 mL) at0° C. The mixture is stirred at the same temperature for 3.5 h, then asolution of ZnBr₂—LiBr in dibutyl ether (2.17 mL of 1.05 M, 2.28 mmol)is added dropwise. The cooling bath is removed and the mixture isstirred at RT for 1 h then a solution of[(2R,3R,4S,5S,6R)-6-bromo-3,4,5-tris(2,2-dimethylpropanoyloxy)tetrahydropyran-2-yl]methyl2,2-dimethylpropanoate (2.00 g, 3.45 mmol) in toluene (3.60 mL) isadded. The final mixture is placed on pre-heated oil bath at 90° C. for4 h (TLC showed lots of starting material), continued at 100° C. for 22h. The resulting mixture is cooled to RT, poured into aqueous 1N HClsolution (40 mL), extracted with EtOAc (3×25 mL). The combined organicextracts are washed with brine, dried over Na₂SO₄, concentrated,purified on Biotage™ SNAP 100 g silica gel cartridge using EtOAc-Hex (0%to 15%, 8 CV) as eluent afforded the title compound (1.00 g, 44%). (Ref.Sebastien Lemaire et. al. Org. Letts. 2012, 14, 1480-1483)

Preparation of Intermediate O:

(2R,3R,4R,5R,6R)-2-(acetoxymethyl)-6-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)tetrahydro-2H-pyran-3,4,5-triylTriacetate

Intermediate I (6.640 g, 13.63 mmol) is dissolved in DMF (100 mL) andthe mixture is degassed 3 times with house vacuum and N₂.4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolane(5.191 g, 20.44 mmol), KOAc (5.351 g, 54.52 mmol) and Pd(dppf)Cl₂.CH₂Cl₂(1.113 g, 1.363 mmol) are added and the mixture is heated to 60° C. for24 hrs. The resulting mixture is cooled down to RT and filtered onCelite. The DMF fraction is washed with Hex (3×100 mL), diluted withEtOAc (300 mL). The organic layer is washed with 100 mL of aqueous NH₄Cl(20%), water (2×100 mL), brine, dried over Na₂SO₄ then evaporate todryness. The residue is purified Biotage™ SNAP 340 g silica gelcartridge using EtOAc-Hex (7 to 60%) as eluent to afford title productcontaminated with dioxaborolane. The residue is triturated in heptane at0° C. and the resulting oil is isolated by decantation and finally driedunder vacuum to afford the title compounds as colorless oil (7.512 g).LC-MS: m/z=557.4 (M+Na⁺).

Preparation of Intermediate P:

((2R,3R,4R,5R,6R)-6-ethynyl-3,4,5-tris((triisopropylsilyl)oxy)tetrahydro-2H-pyran-2-yl)methanol

To a solution of Intermediate L (1.02 g, 1.40 mmol) in MeOH (10 mL) isadded K₂CO₃ (386 mg, 2.80 mmol). After stirring for 1 h, the reaction istreated with prewashed Dowex 50WX4-400 resin, filtered and washed withportions of MeOH. The combined filtrates are concentrated to provide acolorless gum which is purified by flash chromatography on a Biotage™SNAP 25 g cartridge, using a gradient of EtOAc in Hex, 0-20% as eluent.Combined fractions concentrated to provide the title compound as acolorless (878 mg, 96% yield).

Preparation of Intermediate Q:

(2R,3R,4R,5R,6R)-2-(4-bromo-2-methylphenyl)-6-((pivaloyloxy)methyl)tetrahydro-2H-pyran-3,4,5-triyltris(2,2-dimethylpropanoate)

Intermediate Q is prepared according to the procedure described forIntermediate N but using 4-bromo-1-iodo-2-methyl-benzene as startingmaterial.

Preparation of Intermediate R:

(2R,3R,4R,5R,6R)-2-(2-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-6-((pivaloyloxy)methyl)tetrahydro-2H-pyran-3,4,5-triyltris(2,2-dimethylpropanoate)

Intermediate R is prepared according to the procedure described forIntermediate 0 but using Intermediate Q as starting material.

The following is a list of key Intermediates which are used in thepreparation of Compounds described therein:

Preparation of Intermediate AG1

Ethyl 2-(2,7-dibromocarbazol-9-yl)acetate

To a solution of 2,7-dibromo-9H-carbazole (3.00 g, 9.23 mmol) in CH₃CN(30 mL) are added ethyl 2-iodoacetate (5.926 g, 3.274 mL, 27.69 mmol)and Cs₂CO₃ (3.008 g, 9.231 mmol). The reaction mixture is heated toreflux for 4 h, cooled to RT, diluted with EtOAc (50 mL), filteredthrough sintered funnel, solids are washed with EtOAc (20 mL) and CH₂Cl₂(20 mL). The filtrate is washed with aqueous NH₄Cl solution, brine,dried over Na₂SO₄, filtered and concentrated to afford solid. The latteris dissolved in CH₂Cl₂, heptane is added, CH₂Cl₂ is slowly removed undervacuo and the resultant precipitate is filtered, washed with heptane toafford the title compound (1.75 g, 3.97 mmol, 43%) as light yellowsolid. ESI-MS m/z calc. 410.16. found 410.16 (M+1)⁺.

Preparation of Intermediate AG2:

(2R,3S,4R,5S,6R)-2-Ethynyl-6-(hydroxymethyl)tetrahydropyran-3,4,5-triol

To a mixture of ethyl 2-(2,7-dibromocarbazol-9-yl)acetate (500 mg, 1.216mmol) in THF (3.0 mL) is added borane (1.58 mL of 1 M, 1.58 mmol) in THFat RT. The reaction mixture is stirred at RT for 5.5 h (LC-MS showed 80%of the SM), Reaction mixture is continued for 88 h. It is carefullyquenched with THF:Water (1:1, 4 mL) mixture, basified with solid K₂CO₃,extracted with EtOAc (2×10 mL), combined extracts are washed with brine,dried, concentrated to afford the title compound (440 mg, 88.19%) aswhite solid. ¹H NMR (400 MHz, CDCl₃) δ 7.87 (d, J=8.3 Hz, 2H), 7.60 (d,J=1.4 Hz, 2H), 7.34 (dd, J=8.3, 1.5 Hz, 2H), 4.37 (t, J=5.4 Hz, 2H),4.09-4.01 (m, 2H). ESI-MS m/z found 368.18 (M+1)⁺.

Preparation of Intermediate AG3:

2,7-Dibromo-9-[2-(trideuteriomethoxy)ethyl]carbazole

To a cold (0° C.) stirred suspension of 60% NaH in oil (51 mg, 1.27mmol) (prewashed with toluene) in DMF (1 mL) is added a solution ofIntermediate AG3, (260 mg, 0.634 mmol) in DMF (1 mL). The reactionmixture is stirred at 0° C. 45 min, treated withtrideuterio(iodo)methane (118 μL, 1.90 mmol), stirred for 20 min. Thecooling bath is removed, and the mixture is stirred 45 min, quenchedwith aqueous NH₄Cl solution. The resulting precipitate is filtered,washed with water, dried under high vacuum to afford the title compound(235 mg, 78%) as white solid. ¹H NMR (400 MHz, CDCl₃) δ 7.87 (d, J=8.3Hz, 2H), 7.58 (d, J=1.5 Hz, 2H), 7.33 (dd, J=8.3, 1.6 Hz, 2H), 4.37 (t,J=5.7 Hz, 2H), 3.74 (t, J=5.7 Hz, 2H).

Preparation of Intermediate AG4:

2,7-Dibromospiro[fluorene-9,4′-piperidine]

Step I: tert-Butyl2,7-dibromospiro[fluorene-9,4′-piperidine]-1′-carboxylate. IntermediateAG5

To a cold (0° C.) stirred solution of 2,7-dibromo-9H-fluorene (7.500 g,23.15 mmol) in THF (38 mL) is added NaH in oil (3.7 g, 146.5 mmol) inportions. Cooling bath is removed, mixture stirred at RT for 30 min(rapid evolution of hydrogen gas observed) cooled back to 0° C. Asolution of tert-butyl N,N-bis(2-chloroethyl)carbamate (4.00 g, 16.52mmol) in THF (6.0 mL) is then added. The reaction mixture is slowlywarmed to reflux and stirred for 4 h (bath temp. 75° C.). The finalreaction mixture is cooled to RT, quenched slowly by pouring ontocrushed ice, extracted with Et₂O (2×100 mL). The combined organicextracts are washed with brine, dried over Na₂SO₄ and concentrated. Theresidue is purified on Biotage™ Snap silica gel cartridge (340 g) usinga gradient of EtOAc in Hex (0% to 10%, 6 CV and 10%) as eluent to affordthe title compound (5.00 g, 61%) as an orange solid. ¹H NMR (400 MHz,CDCl₃) δ 7.71 (d, J=1.7 Hz, 2H), 7.56 (d, J=8.1 Hz, 2H), 7.49 (dd,J=7.9, 1.5 Hz, 2H), 3.92-3.72 (m, 4H), 1.94-1.74 (m, 4H), 1.53 (s, 9H).

Step II: Intermediate AG4

To solution of HCl in dioxane (39.0 mL of 4 M, 156 mmol) is addedIntermediate AG5, (4.50 g, 9.12 mmol) at RT. The mixture is stirred atRT for 45 min (product precipitated within 5 min), diluted withanhydrous Et₂O (80 mL), cooled to ˜4° C. and filtered. The precipitateis washed with Et₂O (40 mL) and dried to afford the title compound(3.567 g, 84%) as a white solid. ¹H NMR (400 MHz, DMSO-D₆) δ 8.95 (brs,2H), 7.91 (d, J=1.5 Hz, 2H), 7.88 (d, J=8.2 Hz, 2H), 7.62 (dd, J=8.1,1.6 Hz, 2H), 3.50-3.38 (m, 4H), 2.08-1.97 (m, 4H). ESI-MS m/z calc.390.95712. found 394.19 (M+1)⁺.

Preparation of Intermediate AG6

2-[2,7-Dibromo-9-(2-hydroxyethyl)fluoren-9-yl]ethanol

Step I: 2-[2,7-dibromo-9-(2-oxoethyl)fluoren-9-yl]acetaldehyde

To a stirred solution of 2′,7′-dibromospiro[cyclopentene-4,9′-fluorene](500 mg, 1.33 mmol) in CH₂Cl₂ (10 mL) is added MeOH (100 μL, 2.47 mmol)and cooled to −78° C. A stream of O₃/O₂ mixture is bubbled through thesolution until blue color persist (10 min), and then excess ozone isflushed off with nitrogen gas until the solution is clear. The reactionmixture is quenched with methyl sulfide (0.38 mL, 6.85 mmol), stirredfor 1 h, the cooling bath removed, stirred for 1 h, concentrated toafford the title compound. The latter is used without purification inthe next step.

Step II: Intermediate AG6

To a cold (0° C.) stirred solution of2-[2,7-dibromo-9-(2-oxoethyl)fluoren-9-yl]acetaldehyde from Step I (0.66mmol) in a mixture of MeOH (2 mL) and THF (2 mL) is added NaBH₄ (100 mg,2.64 mmol) in one portion, after stirring for 1.5 h, the reactionmixture is quenched with water (5 mL) and aqueous 1N HCl (5 mL). Themixture is concentrated and the resulting aqueous phase is extractedwith EtoAc (2×15 mL). The combined extracts are washed with brine, driedover Na₂SO₄, and concentrated. The residue is purified on Biotage™ SNAPsilica gel cartridge (25 g) using EtOAc in Hex (20% to 75%) as eluent toafford the title compound (230 mg, 84%) as white solid. ¹H NMR (400 MHz,CD₃OD) δ 7.67 (d, J=1.7 Hz, 2H), 7.65 (d, J=8.1 Hz, 2H), 7.50 (dd,J=8.1, 1.7 Hz, 2H), 2.84-2.75 (m, 4H), 2.36-2.27 (m, 4H).

Preparation of Intermediate AG7

(1R,2S)-2′,7′-Dibromospiro[cyclopentane-4,9′-fluorene]-1,2-diol

A 100 mL flask is charged with potassium carbonate (880 mg, 6.367 mmol),hexacyanoiron(3-) (Potassium Ion (3)) (2.1 g, 6.378 mmol),1,4-bis[(S)-[(2R,4S,5R)-5-ethylquinuclidin-2-yl]-(6-methoxy-4-quinolyl)methoxy]phthalazine(17 mg, 0.022 mmol)[(DHQD)₂PHAL] and dipotassium dioxido-dioxo-osmiumdihydrate (2 mg, 0.005 mmol), to this is added water (5.0 mL) andt-Butanol (4.0 mL), stirred for 15 min, methanesulfonamide (304 mg,3.196 mmol) is added, stirred for 15 min. To the yellow-orange mixtureis added a warm solution of2′,7′-dibromospiro[cyclopentene-4,9′-fluorene] (200 mg, 0.5318 mmol) inEtOAc (1.0 mL) and stirred vigorously for 15 h. The reaction mixture isquenched with sodium sulfite (1.4 g, 13.59 mmol), stirred for 1 h,filtered through Celite cartridge, flask is rinsed with EtOAc (4×5 mL),combined filtrate is dried (Na₂SO₄), and concentrated. The residue ispurified on Biotage™ SNAP silica gel cartridge (25 g) eluting with agradient of EtOAc in hexanes (10% to 50%) to afford the title compound(200 mg, 91.7%) as white solid. ¹H NMR (400 MHz, CD₃OD) δ 8.04 (d, J=1.8Hz, 1H), 7.64-7.53 (m, 3H), 7.49-7.38 (m, 2H), 4.45 (t, J=4.5 Hz, 2H),2.31-2.12 (m, 4H). ESI-MS m/z calc. 407.93607. found 408.06 (M+1)⁺.

Preparation of Intermediate AG8

1-(2,7-Dibromospiro[fluorene-9,4′-piperidine]-1′-yl)ethanone

To a stirred mixture of 2,7-dibromospiro[fluorene-9,4′-piperidine](Hydrochloric Acid (1)), AG4, (1000 mg, 2.142 mmol) in DMF (10 mL) isadded Et₃N (900 μL, 6.457 mmol), to the resultant clear solution isadded drop wise acetyl chloride (180 μL, 2.532 mmol) at RT (exotherm).The resultant reaction mixture is stirred for 1 h, diluted with water(˜20 mL)), stirred for 20 min, resultant precipitate is filtered, washedwith water (10 mL), heptane (6 mL), and dried under high vacuum toafford the title compound (885 mg, 94.4%) as white solid. ¹H NMR (400MHz, DMSO-D₆) δ 7.99 (brs, 2H), 7.84 (d, J=8.1 Hz, 2H), 7.57 (brd, J=8.1Hz, 2H), 3.88-3.71 (m, 4H), 2.08 (s, 3H), 1.88-1.78 (m, 2H), 1.76-1.65(m, 2H). ESI-MS m/z calc. 432.96768. found 434.2 (M+1)⁺.

Preparation of Intermediate AG9

1-(2,7-Dibromospiro[fluorene-9,4′-piperidine]-1′-yl)-2-hydroxy-2-methyl-propan-1-one

To a stirred mixture of 2,7-dibromospiro[fluorene-9,4′-piperidine](Hydrochloric Acid (1)), AG4, (60 mg, 0.1397 mmol) and HATU (64 mg,0.1683 mmol) in DMF (1 mL) is added sequentially2-hydroxy-2-methyl-propanoic acid (17.5 mg, 0.168 mmol) and Et₃N (60 μL,0.4305 mmol) at RT, stirred for 2 h, diluted with water (˜3 mL),resultant precipitate is filtered, washed with water, heptane (1 mL),and dried under high vacuum to afford the title compound (52 mg, 68.1%)as white solid. ¹H NMR (400 MHz, CDCl₃) δ 7.71 (brs, J=1.5 Hz, 2H), 7.58(d, J=8.1 Hz, 2H), 7.52 (d, J=8.1 Hz, 2H), 4.37 (s, 1H), 4.12-4.01 (m,4H), 1.94-1.84 (m, 4H), 1.60 (s, 6H).

Preparation of Intermediate A10

Methyl 2,7-dibromospiro[fluorene-9,4′-piperidine]-1′-carboxylate

To a cold (0° C.) stirred mixture of2,7-dibromospiro[fluorene-9,4′-piperidine] (Hydrochloric Acid (1)), AG4,(60 mg, 0.1397 mmol) in CH₂Cl₂ (1 mL) is added methyl chloroformate (16μL, 0.207 mmol) followed by Et₃N (58 μL, 0.419 mmol), stirred for 1.5 h.Reaction mixture is quenched with aqueous 1N HCl (1.5 mL), extractedwith methylene chloride (3×2 mL). The combined filtrate is passedthrough phase separator, and concentrated to afford the title compound(59 mg, 88.74%) as white solid. ¹H NMR (400 MHz, CDCl₃) δ 7.71 (d, J=1.3Hz, 2H), 7.57 (d, J=8.1 Hz, 2H), 7.50 (dd, J=8.1, 1.5 Hz, 2H), 3.87 (s,4H), 3.78 (s, 3H), 1.84 (s, 4H).

Preparation of Intermediate A11

2,7-Dibromo-1′-methylsulfonyl-spiro[fluorene-9,4′-piperidine]

To a cold (0° C.) stirred mixture of2,7-dibromospiro[fluorene-9,4′-piperidine] (Hydrochloric Acid (1)), AG4,(60 mg, 0.1397 mmol) in CH₂Cl₂ (1 mL) is added methane sulfonyl chloride(16 μL, 0.2096 mmol) followed by Et₃N (58 μL, 0.4191 mmol), stirred for1.5 hr (LC-MS showed complete conversion). The reaction mixture isquenched with aqueous 1N HCl (1.5 mL), extracted with methylene chloride(3×2 mL). The combined filtrate is passed through phase separator, andconcentrated to afford the title compound (62 mg, 0.1240 mmol, 88.74%)as white solid. ¹H NMR (400 MHz, CDCl₃) δ 7.69 (d, J=1.5 Hz, 2H), 7.58(d, J=8.1 Hz, 2H), 7.52 (dd, J=8.1, 1.6 Hz, 2H), 3.71-3.63 (m, 4H), 2.99(s, 3H), 2.04-1.95 (m, 4H).

Preparation of Intermediate AG12

[9-(Acetoxymethyl)-2,7-dibromo-fluoren-9-yl]methyl Acetate

Step I: [2,7-dibromo-9-(hydroxymethyl)fluoren-9-yl]methanol

To a mixture of [9-(hydroxymethyl)fluoren-9-yl]methanol (1.584 g, 7mmol) in water (5.5 mL) is added conc. H₂SO₄ (3 drops) and heated at 50°C. for 4.5 h, cooled to RT, quenched with aqueous sodium thiosulfate,diluted with water (30 mL), filtered, precipitate is washed with water(15 mL), and dried under high vacuum to afford the title compound (2.5g, 70.3%) as white solid. The product contains ˜15% of unknown impurityand it has been used as such in the next step without furtherpurification. In LC-MS, major peak appeared at 349 (M−2×H₂O)⁺

Step II

To a stirred mixture of[2,7-dibromo-9-(hydroxymethyl)fluoren-9-yl]methanol (250 mg, 0.492 mmol)in CH₂Cl₂ (5 mL) and pyridine (1 mL) is added 4-dimethylaminopyridine(25 mg, 0.205 mmol) and acetic anhydride (1 mL, 10.6 mmol) at RT, after1.5 h, quenched with aqueous 1 N HCl (3 mL), stirred for 30 min,extracted with methylene chloride (3×5 mL), combined extracts are washedwith aqueous 1 N HCl (2×4 mL), passed through phase separator, dried(Na₂SO₄), and concentrated. The residue is purified on Biotage™) SNAPsilica gel cartridge (50 g) eluting with a gradient of EtOAc in Hex (10%to 40%, 8 CV) to afford the title compound (120 mg, 52.11%) as whitesolid. A peak at 349 appeared in the LC-MS (M−2×OAc). ¹H NMR (400 MHz,CDCl₃) δ 7.71-7.67 (m, 2H), 7.58-7.55 (m, 4H), 4.29 (s, 4H), 2.10 (s,6H).

Preparation of Intermediate AG13

2,7-Dibromospiro[fluorene-9,5′-oxepane]-2′-one

To a stirred solution of2′,7′-dibromospiro[cyclohexane-4,9′-fluorene]-1-one (277 mg, 0.6821mmol) in CH₂Cl₂ (7 mL) is added m-CPBA (155 mg, 0.8982 mmol). Thereaction mixture is stirred at RT for 18 h, aqueous saturated NaHCO₃ isadded, extracted with CH₂Cl₂. The organic phases are combined, driedover MgSO₄, filtered, and concentrated. The residue is purified onBiotage™ SNAP silica gel cartridge with a gradient of EtOAc in Hex(0-20%, 20 CV) to afford the title compound (205 mg, 71.2%) as a whitesolid.

Preparation of Intermediate AG14

2,7-Dibromo-1′-methyl-spiro[fluorene-9,4′-piperidine]

To a stirred mixture of 2,7-dibromospiro[fluorene-9,4′-piperidine](Hydrochloric Acid (1)), AG4, (100 mg, 0.233 mmol) in 1,2-dichloroethane(2 mL) is added aqueous formaldehyde (38 μL of 37% w/v, 0.4683 mmol)followed by triacetoxyboranuide (sodium ion (1)) (148.0 mg, 0.698 mmol)in one portion at RT. Reaction mixture is stirred at RT for 1 h (LC-Msshowed the presence of clean product). It is quenched with aqueous 10%NaOH until pH 6-7, extracted with methylene chloride (4×5 mL), combinedextracts are dried (Na₂SO₄) and concentrated to afford the titlecompound (91 mg, 93.9%). ¹H NMR (400 MHz, CDCl₃) δ 7.79 (d, J=1.3 Hz,2H), 7.54 (d, J=8.1 Hz, 2H), 7.48 (dd, J=8.1, 1.7 Hz, 2H), 2.91-2.79 (m,4H), 2.54 (s, 3H), 2.02-1.89 (m, 4H). ESI-MS m/z calc. 404.97278, found406.28 (M+1)⁺.

Preparation of Intermediate AG15

2,7-dibromo-9-propyl-carbazole

To a solution of 2,7-dibromo-9H-carbazole (150 mg, 0.462 mmol) in MeCN(9 mL) is added 1-iodopropane (135 μL, 1.38 mmol) and Cs₂CO₃ (752 mg,2.31 mmol). The reaction is stirred for 18h at reflux. The resultingmixture is cooled to RT, filtered and diluted with EA. The organic phaseis washed with saturated aqueous NH₄Cl, dried over MgSO₄, andconcentrated. The residue is purified on Biotage™ SNAP silica gelcartridge (0-5% EA/Hex) to give 2,7-dibromo-9-propyl-carbazole (145 mg,86%) as a white solid.

Intermediates AG16, AG24, AG25 are prepared according to the proceduredescribed for Intermediate AG15 using the appropriated alkylatingreagent.

Intermediate AG16:

2,7-Dibromo-9-methyl-carbazole

¹H NMR (400 MHz, Chloroform-d) δ 7.88 (d, J=8.3 Hz, 2H), 7.53 (d, J=1.6Hz, 2H), 7.33 (dd, J=8.3, 1.7 Hz, 2H), 3.77 (s, 3H).

Intermediate AG24:

2-(2,7-Dibromocarbazol-9-yl)-1-morpholino-ethanone

¹H NMR (400 MHz, Chloroform-d) δ 7.88 (d, J=8.3 Hz, 2H), 7.40 (s, 2H),7.35 (dd, J=8.3, 1.4 Hz, 2H), 4.96 (s, 2H), 3.77-3.49 (m, 8H).

Intermediate AG25:

2-(2,7-Dibromocarbazol-9-yl)-N,N-dimethyl-acetamide

¹H NMR (400 MHz, Chloroform-d) δ 7.88 (d, J=8.3 Hz, 2H), 7.41 (s, 2H),7.34 (d, J=8.3 Hz, 2H), 4.96 (s, 2H), 3.17 (s, 3H), 3.02 (s, 3H).

Preparation of Intermediate AG17

2,7-dibromo-9-pentyl-fluoren-9-ol

A solution of 2,7-dibromofluoren-9-one (200 mg, 0.592 mmol) in THF (2mL) is added to a solution of n-pentyl magnesium bromide in Et₂O (355 μLof 2.0 M, 0.710 mmol) diluted with Et₂O (4 mL) at 0° C. The reaction isstirred 18h at RT and quenched with 1N Na₂CO₃. The organic phase isseparated, washed with brine, dried over MgSO₄, filtered andconcentrated. The residue is purified on Biotage™ SNAP silica gelcartridge (0-8% EA/hex) to give 2,7-dibromo-9-pentyl-fluoren-9-ol (150mg, 62%) as a white solid.

Intermediates AG18, AG19, AG20, AG21, AG22, and AG23 are preparedaccording to the procedure described for Intermediate AG17 using theappropriated Grignard reagent.

Intermediate Name/Structure ¹HNMR AG182,7-Dibromo-9-cyclopropyl-fluoren- (400 MHz, Chloroform-d) δ 7.65 (d, J= 9-ol 8.0 Hz, 2H), 7.52-7.39 (m, 4H), 1.90

(s, 1H), 1.15-1.00 (m, 1H), 0.79-0.66 (m, 2H), 0.51 (dd, J = 10.2, 4.2Hz, 2H). AG19 2,7-Dibromo-9-isopropyl-fluoren-ol (400 MHz, Chloroform-d)δ 7.62 (d, J =

1.5 Hz, 2H), 7.48 (dd. J = 8.0, 1.7 Hz, 2H), 7.43 (d, J = 8.1 Hz, 2H),2.43 (p, J = 6.9 Hz, 1H), 2.03 (s, 1H), 0.81 (d, J = 6.8 Hz, 6H). AG202,7-Dibroino-9-ethyl-fluoren-9-ol (400 MHz, Chloroform-d) δ 7.61 (d, J =

1.7 Hz, 2H), 7.48 (dd, J = 8.0, 1.7 Hz, 2H), 7.44 (d, J = 8.1 Hz, 2H),2.14 (q, J = 7.5 Hz, 2H), 2.02 (s, 1H), 0.54 (t, J = 7.5 Hz, 3H). AG212,7-Dibromo-9-propyl-fluoren-9-ol (400 MHz, Chloroform-d) δ 7.62 (d, J =

1.7 Hz, 2H), 7.48 (dd, J = 8.0, 1.7 Hz, 2H), 7.43 (d, J = 8.0 Hz, 2H),2.11- 2.03 (m, 2H), 2.01 (s, 1H), 0.97-0.84 (m, 2H), 0.78 (t, J = 7.0Hz, 3H). AG22 2,7-Dibromo-9-isobutyl-fluoren-9-ol (400 MHz,Chloroform-d) δ 7.62 (d, J =

1.7 Hz, 2H), 7.49 (dd, J = 8.1, 1.8 Hz, 2H), 7.44 (d, J = 8.0 Hz, 2H),2.10 (d, J = 6.3 Hz, 2H), 1.95 (s, 1H), 1.53 (s, 2H), 1.17 (dt, J =13.0, 6.2 Hz, 1H), 0.59 (d, J = 6.7 Hz, 6H). AG23 2,7-Dibromo-9-(3- (400MHz, Chloroform-d) δ 7.48 (d, J = methoxyphenyl)fluoren-9-ol 1.0 Hz,4H), 7.45-7.41 (m, 2H), 7.19 (t,

J = 8.0 Hz, 1H), 7.00-6.96 (m, 1H), 6.84-6.77 (m, 2H), 3.77 (s, 3H),2.42 (s, 1H).Preparation of Intermediate AG26

3-(2,7-dibromo-9-hydroxy-fluoren-9-yl)propane-1,2-diol

Step I: 9-Allyl-2,7-dibromo-fluoren-9-ol

The title compound is prepared according to the procedure described forIntermediate AG17 using ally magnesium bromide. ¹H NMR (400 MHz,Chloroform-d) δ 7.64 (d, J=1.7 Hz, 2H), 7.51-7.40 (m, 4H), 5.62-5.48 (m,1H), 5.05-4.95 (m, 2H), 2.77 (d, J=7.3 Hz, 2H), 2.18 (s, 1H).

Step II: Intermediate AG26

To a solution of 9-allyl-2,7-dibromo-fluoren-9-ol from Step I (250 mg,0.658 mmol) in acetone/H₂O 4:1 (6.6 mL) were added4-methyl-4-oxido-morpholin-4-ium (193 mg, 1.65 mmol) and OsO₄ 2.5% intBuOH (401 μL of 2.5% w/v, 0.03943 mmol). The mixture is stirred at RTfor 18 h. The reaction is quenched with the addition of saturatedaqueous NH₄Cl. The mixture is extracted with EtOAc. The organic phase iswashed with saturated aqueous NH₄Cl, dried over MgSO₄, filtered andconcentrated. The residue is purified over Biotage™ SNAP silica gelcartridge (30-100% EA/hex 20 CV) to give the title compound (174 mg,64%) as a white solid. ¹H NMR (400 MHz, Chloroform-d) δ 7.82 (s, 1H),7.67 (s, 1H), 7.54-7.40 (m, 4H), 4.26-4.13 (m, 1H), 3.76-3.67 (m, 1H),3.66-3.55 (m, 1H), 3.52-3.37 (m, 2H), 2.49-2.37 (m, 1H), 1.86 (d, J=5.4Hz, 2H), 1.59 (dd, J=8.1, 3.5 Hz, 1H).

Preparation of Intermediates AG27

(3'S,4′R)-2,7-Dibromospiro[fluorene-9,6′-tetrahydropyran]-3′,4′-diol and(3′R,4′S)-2,7-dibromospiro[fluorene-9,6′-tetrahydropyran]-3′,4′-diol

Step I: 9-Allyl-9-allyloxy-2,7-dibromo-fluorene

To a suspension of NaH (60% suspended in oil, 526 mg, 13.15 mmol) in DMF(15 mL) at 0° C. was added a solution of9-allyl-2,7-dibromo-fluoren-9-ol from Intermediate AG26 Step I (1.00 g,2.63 mmol) in DMF (7.5 mL). The reaction is warmed to RT for 1 h, cooledto 0° C. and allyl bromide (797 μL, 9.210 mmol) is added. The reactionmixture is stirred at RT for 18 h. The resulting mixture is diluted withEtOAc and quenched with saturated aqueous NH₄Cl. The organic phase isseparated, dried over MgSO₄, filtered and concentrated. The residue ispurified on Biotage™ SNAP silica gel cartridge (0-8% EA/hex 20 CV) togive the title compound (1.04 g, 94%) as a yellow solid. ¹H NMR (400MHz, Chloroform-d) δ 7.60 (d, J=1.7 Hz, 2H), 7.49 (dd, J=8.1, 1.8 Hz,2H), 7.45 (d, J=8.0 Hz, 2H), 5.74 (ddt, J=17.1, 10.6, 5.4 Hz, 1H), 5.45(ddt, J=17.4, 10.3, 7.2 Hz, 1H), 5.16 (dq, J=17.2, 1.7 Hz, 1H), 5.05(dq, J=10.5, 1.4 Hz, 1H), 4.91-4.81 (m, 2H), 3.36 (dt, J=5.4, 1.5 Hz,2H), 2.78 (d, J=7.2 Hz, 2H).

Step II: 2′,7′-Dibromospiro[2,5-dihydropyran-6,9′-fluorene]

To a solution of 9-allyl-9-allyloxy-2,7-dibromo-fluorene from Step I(1.04 g, 2.48 mmol) in CH₂Cl₂ (200 mL) at RT is added Grubbs 2^(nd) gen.catalyst (105 mg, 0.124 mmol). The reaction mixture is stirred at RT for18 h. The resulting mixture is concentrated and the residue is purifiedon Biotage™ SNAP silica gel cartridge (0-8% EA/hex 20 CV) to give thetitle compound (853 mg, 88%) as a white solid. ¹H NMR (400 MHz,Chloroform-d) δ 7.68 (d, J=1.4 Hz, 2H), 7.53-7.44 (m, 4H), 6.18-6.02 (m,1H), 4.54-4.41 (m, 2H), 2.53 (s, 2H).

Step III: Intermediates AG27

The title compounds are prepared using2′,7′-Dibromospiro[2,5-dihydropyran-6,9′-fluorene] from Step II (250 mg,0.6376 mmol) as starting material according to the procedure describedin Compound 196 Step II to give the title racemic mixture ofdiastereoisomers as a white solid (201 mg, 74% yield). ¹H NMR (400 MHz,Chloroform-d) δ 7.83 (s, 1H), 7.68 (s, 1H), 7.57-7.41 (m, 4H), 4.54-4.43(m, 1H), 4.20 (d, J=13.3 Hz, 1H), 4.13 (d, J=8.6 Hz, 2H), 2.46-2.33 (m,2H), 2.27 (dd, J=7.1, 1.8 Hz, 1H), 2.00-1.91 (m, 1H).

Preparation of Intermediate AG28

1-(2,7-Dibromo-9H-fluoren-9-yl)-4-methyl-piperazine

To a solution of 2,7-dibromo-9H-fluoren-9-ol (250 mg, 0.735 mmol) inCH₂Cl₂ (3 mL) and THF (1 mL) is added thionyl chloride (120 μL, 1.65mmol). The mixture is stirred at RT for 1 h. The resulting mixture isconcentrated to dryness. MeCN (3 mL) is added then 1-methylpiperazine(408 μL, 3.67 mmol) is added and the mixture is stirred at 82° C. for 18h. The reaction mixture is quenched by addition of saturated aqueousNH₄Cl and diluted with EtOAc. The organic phase is separated, washedwith saturated aqueous NH₄Cl, dried over MgSO₄, filtered andconcentrated. The residue is purified over Biotage™ SNAP silica gelcartridge (30-100% EA/hex+1% Et₃N buffer 20 CV) to give the titlecompound (200 mg, 64% yield) as a yellow solid. ¹H NMR (400 MHz,Chloroform-d) δ 7.75 (s, 2H), 7.47 (s, 4H), 4.77 (s, 1H), 2.64 (t, J=4.1Hz, 4H), 2.40 (s, 4H), 2.26 (s, 3H).

Preparation of Intermediate AG29

2,7-Dibromospiro[fluorene-9,2′-tetrahydropyran]

To a solution of 2′,7′-dibromospiro[2,5-dihydropyran-6,9′-fluorene] fromIntermediate AG27 Step II (200 mg, 0.510 mmol) in EtOAc (7 mL) is addedSiliaCat Pd⁰ (26 mg, 0.0013 mmol). H₂ is then bubbled for 5 minutes andthe reaction is stirred under H₂ atmosphere for 1 week. The resultingmixture is filtered and concentrated. The residue is purified onBiotage™ SNAP silica gel cartridge (0-8% EA/hex 17 CV) to give the titlecompound (120 mg, 60%) as a pale yellow solid. ¹H NMR (400 MHz,Chloroform-d) δ 7.82 (d, J=1.6 Hz, 2H), 7.51-7.46 (m, 4H), 4.18-4.00 (m,2H), 2.11-2.03 (m, 2H), 1.93 (dt, J=13.3, 6.4 Hz, 4H).

Preparation of Intermediate AG30

N-(2,7-Dibromo-9H-fluoren-9-yl)-N,N,N-trimethyl-ethane-1,2-diamine

The title compound is prepared according to the procedure described forIntermediate A28 but using N1,N1,N2-trimethylethane-1,2-diamine asstarting material. ¹H NMR (400 MHz, Chloroform-d) δ 7.73 (s, 2H), 7.48(s, 4H), 4.87 (s, 1H), 2.74 (t, J=6.3 Hz, 2H), 2.59 (t, J=6.2 Hz, 2H),2.37 (s, 6H), 2.26 (s, 3H).

Preparation of Intermediate AG31

2,7-Dibromo-9-(cyclopentylmethyl)fluoren-9-ol

A Et₂O washed sealed tube under N₂ flow is charged withiodomethylcyclopentane (145 μL) in Et₂O (1 mL) at RT. Magnesium (27 mg,1.111 mmol) and iodine (0.30 mg, 0.0012 mmol) are added and the mixtureis stirred at reflux for 2 h (or until all magnesium was consumed). Theresulting mixture is cooled down to 0° C. and a solution of2,7-dibromofluoren-9-one (250 mg, 0.740 mmol) in THF (2 mL) is added.The reaction is stirred at RT for 18 h. The resultant mixture isquenched with saturated aqueous NaHCO₃, the organic phase is separated,washed with brine, dried over MgSO₄, filtered and concentrated. Theresidue is purified Biotage™ SNAP silica gel cartridge (0-10% EA/hex 20CV) to give the title compound (62 mg, 13%) as a yellow solid. ¹H NMR(400 MHz, Chloroform-d) δ 7.61 (d, J=1.8 Hz, 2H), 7.53-7.40 (m, 4H),2.26 (d, J=5.9 Hz, 2H), 1.98 (s, 1H), 1.41 (s, 2H), 1.27-1.14 (m, 5H),0.82 (s, 2H).

Preparation of Intermediate AG32

2,7-Dibromo-9-(cyclohexylmethyl)fluoren-9-ol

The tile compound is prepared according to the procedure described forIntermediate AG32 but using iodomethylcyclohexane as starting material.¹H NMR (400 MHz, Chloroform-d) δ 7.61 (d, J=1.7 Hz, 2H), 7.48 (dd,J=8.0, 1.8 Hz, 2H), 7.43 (d, J=8.0 Hz, 2H), 2.07 (d, J=5.4 Hz, 2H), 1.94(s, 1H), 1.46 (dd, J=9.0, 3.2 Hz, 3H), 1.22 (d, J=11.8 Hz, 2H),1.05-0.69 (m, 6H).

Preparation of Intermediate A33

2,7-Dibromo-9-(2-hydroxyethyl)fluoren-9-ol

To a solution of 9-allyl-2,7-dibromo-fluoren-9-ol from Intermediate AG26Step I (1.00 g, 2.63 mmol) in CH₂Cl₂ (13 mL) at −78° C. is bubbled O₃until the solution became blue. Nitrogen is then bubbled for 10 minutesto remove excess of O₃. The reaction is allowed to warm at 0° C. andNaBH₄ (597 mg, 15.8 mmol) is added and the mixture is stirred 1. Thereaction mixture is quenched by adding saturated aqueous NH₄Cl. Theorganic phase is separated, washed with saturated aqueous NH₄Cl, driedover MgSO₄, filtered and concentrated. The residue is purified onBiotage™ SNAP silica gel cartridge (0-20% EA/hex 20 CV) to give thetitle compound (443 mg, 44%) as a white solid. ¹H NMR (400 MHz,Chloroform-d) δ 7.72 (dd, J=2.6, 1.5 Hz, 2H), 7.50 (ddd, J=8.1, 3.0, 1.6Hz, 2H), 7.47-7.42 (m, 2H), 3.84 (d, J=3.9 Hz, 2H), 3.27 (s, 1H), 2.21(td, J=5.9, 3.0 Hz, 2H), 2.13 (td, J=4.9, 2.4 Hz, 1H).

Preparation of Intermediate AG34

2,7-dibromo-9-(tetrahydro-2H-pyran-4-yl)-9H-carbazole

To a solution of 2,7-dibromo-9H-carbazole (250 mg, 0.77 mmol) andtetrahydropyran-4-ol (146 μL, 1.54 mmol) in Toluene 2.5 mL is added2-tributylphosphoranylideneacetonitrile (1.54 mL of 1 M, 1.54 mmol). Thereaction mixture is stirred at 110° C. for 4h, cooled down to RT andpartitioned between EtOAc and water. The organic layer is dried overNa₂SO₄, filtered, concentrated and purified Biotage™ SNAP silica gelcartridge (24 g) eluting with 0-50% EtOAC in Hex to afford the tittlecompound (95 mg, 30%). ¹H NMR (400 MHz, Chloroform-d) δ 7.89 (d, J=8.3Hz, 2H), 7.70 (d, J=1.6 Hz, 2H), 7.34 (dd, J=8.3, 1.6 Hz, 2H), 4.58(ddt, J=12.5, 8.8, 4.4 Hz, 1H), 4.24 (dd, J=11.7, 4.8 Hz, 2H), 3.64 (td,J=12.1, 2.0 Hz, 2H), 2.69 (qd, J=12.5, 4.7 Hz, 2H), 1.84 (ddd, J=12.9,4.3, 1.8 Hz, 2H).

Intermediates AG35-40 are prepared according to the procedure describedin Intermediate AG34

Intermediate Name/Structure ¹HNMR AG35 tert-butyl 4-(2,7-dibromo-9H-(400 MHz, Chloroform-d) δ 7.89 (d, carbazol-9-yl)piperidine-1- J = 8.3Hz, 2H), 7.62 (d, J = 1.6 Hz, 2H), carboxylate 7.33 (dd, J = 8.3, 1.6Hz, 2H), 4.48 (tt,

J = 12.6, 4.2 Hz, 1H), 4.40 (m, 2H), 2.93 (t, J = 12.8 Hz, 2H), 2.49(qd, J = 12.5, 4.6 Hz, 2H), 1.88 (d, J = 12.4 Hz, 2H), 1.54 (s, 9H).AG36 2,7-dibromo-9-((3-methyloxetan-3- (400 MHz, Chloroform-d) δ 7.97yl)methyl)-9H-carbazole 7.82 (m, 2H), 7.52 (d, J = 1.6 Hz, 2H),

7.36 (dd, J = 8.3, 1.6 Hz, 2H), 4.71 (d, J = 6.1 Hz, 2H), 4.47-4.27 (m,4H), 1.44 (s, 3H). AG37 2,7-dibromo-9-(1-methylpiperidin-4- (400 MHz,CD₃OD) δ 8.02-7.77 (m, yl)-9H-carbazole 4H), 7.29 (dd, J = 8.4, 1.6 Hz,2H),

4.61 (tt, J = 11.2, 4.0 Hz, 1H), 3.06 (d, J = 11.8 Hz, 2H), 2.66 (qd, J= 12.7, 4.1 Hz, 2H), 2.39 (s, 3H), 2.34 (dd, J = 12.3, 2.5 Hz, 2H), 1.81(ddd, J = 12.6, 4.9, 2.2 Hz, 2H). AG382,7-dibromo-9-((1-methylpiperidin-4- (400 MHz, Chloroform-d) δ 7.87 (dd,yl)methyl)-9H-carbazole J = 8.3, 0.5 Hz, 2H), 7.49 (d, J = 1.6 Hz,

2H), 7.32 (dd, J = 8.3, 1.6 Hz, 2H), 4.18 (ddd, J = 7.9, 6.9, 1.3 Hz,2H), 2.77-2.67 (m, 1H), 2.55 (ddd, J = 9.2, 7.7.6.1 Hz, 1H), 2.47 (td, J= 8.8, 5.7 Hz, 1H), 2.25 (s, 3H), 2.17 (td, J = 6.2, 4.2 Hz, 2H),2.00-1.77 (m, 2H), 1.45 (ddt, J = 12.0, 8.1, 5.7 Hz, 1H). AG394-(2-(2,7-dibromo-9H-carbazol-9- (400 MHz, Chloroform-d) δ 7.87 (dd,yl)ethyl)morpholine J = 8.3, 3.0 Hz, 2H), 7.61-7.49 (m,

2H), 7.33 (dt, J = 8.3, 1.7 Hz, 2H), 4.31 (td, J = 7.8, 7.3, 3.4 Hz,2H), 3.68 (t, J = 4.6 Hz, 4H), 2.72 (td, J = 7.1, 2.4 Hz, 2H), 2.61 2.42(m, 4H). AG40 1-(2-(2,7-dibromo-9H-carbazol-9- (400 MHz, Chloroform-d) δ7.88 (d, yl)ethyl)pyrrolidin-2-one J = 8.2 Hz, 2H), 7.57 (d, J = 1.6 Hz,2H),

7.35 (dd, J = 8.3, 1.6 Hz, 2H), 4.47 (t, J = 5.7 Hz, 2H), 3.64 (t, J =5.7 Hz, 2H), 2.64 (t, J = 7.0 Hz, 2H), 2.27 (t, J = 8.1 Hz, 2H), 1.57(t, J = 8.1 Hz, 2H).Preparation of Intermediate AG41

2′,7′-dibromospiro[cyclohexane-1,9′-fluoren]-4-one

Step I: (1,3-dioxolane-2,2-diyl)bis(ethane-2,1-diyl) dimethanesulfonate

To a cold (0° C.) solution of2-[2-(2-hydroxyethyl)-1,3-dioxolan-2-yl]ethanol (1000 mg, 6.200 mmol) inCH₂Cl₂ (10.0 mL) is added Et₃N (2.15 mL, 15.4 mmol) and methanesulfonylchloride (1.00 mL, 12.9 mmol) dropwise over 10 min. The reaction mixtureis stirred 45 min at 0° C., quenched with water (20 mL). The organicphase is washed with saturated aqueous NaHCO₃, dried over Na₂SO₄,filtered and concentrated to afford the title compound (1750 mg, 89%) aswhite solid.

Step II:2,7-dibromodispiro[fluorene-9,1′-cyclohexane-4′,2″-[1,3]dioxolane]

To a cold (0° C.) of 2,7-dibromo-9H-fluorene (1.020 g, 3.100 mmol) inTHF (7.0 mL) is added NaH (60%) in oil (814 mg, 20.4 mmol) in 3portions. The reaction mixture is stirred at RT for 30 min, cooled backto 0° C. and a solution of (1,3-dioxolane-2,2-diyl)bis(ethane-2,1-diyl)dimethanesulfonate from Step I (700 mg, 2.2 mmol) in THF (3.5 mL) isadded. The reaction mixture is stirred at 75° C. for 4h, cooled to RT,quenched with ice-water, extracted with EtOAc (3×10 mL). The combinedextracts are washed with brine, dried over Na₂SO₄ and concentrated.Purification of the residue on Biotage™ SNAP silica gel cartridge (24 g)using an isocratic 50% CH₂Cl₂/Hexane as eluent to afford the tittlecompound (750 mg, 76%). ¹H NMR (400 MHz, Chloroform-d) δ 7.75 (d, J=1.7Hz, 2H), 7.54 (d, J=8.1 Hz, 2H), 7.47 (dd, J=8.1, 1.7 Hz, 2H), 4.11-4.05(s, 4H), 2.06 (m, 4H), 1.98-1.90 (m, 4H).

Step III: Intermediate AG41

To a solution of2,7-dibromodispiro[fluorene-9,1′-cyclohexane-4′,2″-[1,3]dioxolane] (1400mg, 3.11 mmol) in THF (7.0 mL) is added HCl (5.2 mL of 3 M, 15.6 mmol).The reaction mixture is stirred at 45° C. for 16 h, cooled down to RT,diluted with water and extracted with EtOAc. The organic phase isseparated, dried over Na₂SO₄, filtered and concentrated. The residue ispurified on Biotage™ SNAP silica gel cartridge (24 g) eluting withHex/EtOAc (0-50%, 15 CV) to afford the title compound (923 mg, 73%). ¹HNMR (400 MHz, Chloroform-d) δ 7.68 (dd, J=1.7, 0.5 Hz, 2H), 7.60 (dd,J=8.1, 0.5 Hz, 2H), 7.53 (dd, J=8.1, 1.7 Hz, 2H), 2.80 (t, J=7.0 Hz,4H), 2.18 (dd, J=7.4, 6.5 Hz, 4H).

Preparation of Intermediate AG42

2′,7′-dibromospiro[cyclohexane-1,9′-fluoren]-4-ol

To a cold solution (−40° C.) of Intermediate AG41 (75 mg, 0.18 mmol) inTHF (1.5 mL) is added NaBH₄ (10.0 mg, 0.260 mmol). The reaction mixtureis stirred 1h at −40° C. and 2h at RT. The reception is quenched withwater and extracted with EtOAc. The organic phase is separated, driedover Na₂SO₄, filtered and concentrated. The residue is purified onBiotage™ SNAP silica gel cartridge (12 g) eluting with Hex/EtOAc (0-50%,15 CV) to afford the title compound (55 mg, 73%). ¹H NMR (400 MHz,Chloroform-d) δ 7.96-7.81 (m, 1H), 7.61-7.51 (m, 3H), 7.48 (ddd, J=12.1,8.1, 1.7 Hz, 2H), 4.19-4.00 (m, 1H), 2.13 (dq, J=13.9, 4.7 Hz, 2H),2.01-1.88 (m, 2H), 1.83 (dt, J=9.9, 4.7 Hz, 4H).

Preparation of Intermediate AG43

2′,7′-dibromo-4-methylspiro[cyclohexane-1,9′-fluoren]-4-ol

To a cold solution (−40° C.) of Intermediate AG41 (75 mg, 0.18 mmol) inTHF (7.5 mL) is added methyllithium (140 μL of 1.6 M, 0.22 mmol). Thereaction mixture is stirred 1h at −40° C. and 2h at RT, quenched withwater and extracted with EtOAc. The organic phase is separated, driedover Na₂SO₄, filtered and concentrated. The residue is purified onBiotage™ SNAP silica gel cartridge (12 g) eluting with Hex/EtOAc (0-50%,15 CV) to afford the title compound (25 mg, 32%). ¹H NMR (400 MHz,Chloroform-d) δ 7.68 (dd, J=9.5, 1.8 Hz, 2H), 7.61-7.39 (m, 4H),2.29-2.12 (m, 2H), 2.06-1.93 (m, 2H), 1.93-1.80 (m, 2H), 1.47 (s, 3H).

Preparation of Intermediate AG44

2′,7′-dibromospiro[cyclohexane-1,9′-fluoren]-4-amine

To a stirred solution of Intermediate AG41 (200 mg, 0.490 mmol) andammonium acetate (380 mg, 4.90 mmol) in MeOH (2.0 mL) is added NaBH₃CN(22 mg, 0.34 mmol). The reaction mixture is stirred at RT for 16h,quenched with 1N NaOH (1 mL), extracted with CH₂Cl₂, dried over Na₂SO₄,filtered and concentrated. The residue is purified on Biotage™ SNAPsilica gel cartridge (10 g) eluting with CH₂CL₂/MeOH (0-10%) to affordthe title compound (80 mg, 40%). ¹H NMR (400 MHz, Chloroform-d) δ 7.93(d, J=1.7 Hz, 1H), 7.63-7.37 (m, 5H), 3.16-2.97 (m, 1H), 2.09-1.89 (m,4H), 1.86-1.71 (m, 2H), 1.62 (d, J=13.4 Hz, 2H).

Example 1: Preparation of Compound 1 (Method A)((2R,3S,4R,5S,6R)-2-(Hydroxymethyl)-6-[4-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6(hydroxymethyl)tetrahydropyran-2-yl]phenyl]tetrahydropyran-3,4,5-triol)

Step I:[(2R,3S,6S)-3-Acetoxy-6-[4-[(2R,3S,6S)-3-acetoxy-2-(acetoxymethyl)-3,6-dihydro-2H-pyran-6-yl]phenyl]-3,6-dihydro-2H-pyran-2-yl]methylAcetate

A solution of [(2R,3S,4R)-3,4-diacetoxy-3,4-dihydro-2H-pyran-2-yl]methylacetate (1.642 g, 6.033 mmol) and (4-boronophenyl)boronic acid (1.0 g,6.03 mmol) in CH₃CN (30 mL) is degassed by bubbling N₂ (g) for 5 min.Pd(OAc)₂ (338.6 mg, 1.50 mmol) is added and the reaction mixture isstirred at 40° C. for 5 h. Another portion of Pd(OAc)₂ (338.6 mg, 1.50mmol) is added and heating is continued for a further 15 h. The solventis evaporated and the mixture is diluted with EtOAc (25 mL) and filteredthrough celite. The filtrate is washed with sat NaHCO₃ (2×25 mL), driedover Na₂SO₄ and the solvent is evaporated. The crude product is purifiedon Biotage™ SNAP 50 g silica gel cartridge with a gradient elution of10%-30% EtOAc in Hex and a flow rate of 40 mL/min over 35 min to affordthe title compound (550 mg, 18.14%) as an oil. LC-MS: m/z=525.0 (M+Na).

Step II:[(2R,3S,4R,5S,6R)-3-Acetoxy-6-[4-[(2R,3S,4R,5S,6R)-5-acetoxy-6-(acetoxymethyl)-3,4-dihydroxy-tetrahydropyran-2-yl]phenyl]-4,5-dihydroxy-tetrahydropyran-2-yl]methylAcetate

To a solution of[(2R,3S,6S)-3-acetoxy-6-[4-[(2R,3S,6S)-3-acetoxy-2-(acetoxymethyl)-3,6-dihydro-2H-pyran-6-yl]phenyl]-3,6-dihydro-2H-pyran-2-yl]methylacetate (550 mg, 1.095 mmol) in water (1.6 mL)/THF (9.4 mL) is addedmethanesulfonamide (312.5 mg, 3.285 mmol), OsO₄ (1.1 mL of 2.5% w/v int-BuOH, 0.1095 mmol) and NMO (1.283 g, 10.95 mmol) and the reactionmixture is stirred at RT for 5 days. The solvent is evaporated and thecrude mixture is diluted with a dilute solution of sodium bisulfite (50mL) and after 15 min it is extracted with EtOAc (3×25 mL). The combinedorganic extracts are dried over Na₂SO₄ and the solvent is evaporated.The crude product is purified on Biotage™ SNAP 25 g silica gel cartridgewith a gradient of 5%-10% MeOH in CH₂Cl₂ as the eluent and a flow rateof 24 mL/min over 20 min. The isolated product is dissolved in a minimumamount of MeOH and diluted with Et₂O to precipitate the product. Themixture is filtered and washed with diethyl ether to afford titlecompound (145 mg, 23.21%) as a white solid. LC-MS: m/z=571.4 (M+H⁺)

Step III: Compound 1

[(2R,3S,4R,5S,6R)-3-acetoxy-6-[4-[(2R,3S,4R,5S,6R)-5-acetoxy-6-(acetoxymethyl)-3,4-dihydroxy-tetrahydropyran-2-yl]phenyl]-4,5-dihydroxy-tetrahydropyran-2-yl]methylacetate (145 mg, 0.2541 mmol) is dissolved in MeOH (3 mL) and MeONa inMeOH (47 μL of 25% w/v, 0.22 mmol) is added and the reaction mixture isstirred at RT for 6h. The reaction is neutralized by the addition ofAmberlite IR120H resin until the pH changed to neutral. The reactionmixture is filtered and the filtrate is evaporated to afford the titleproduct (108 mg, 0.258 mmol, 24%) as a white solid. ¹H NMR (400 MHz,CD₃OD) δ 7.51 (s, 4H), 4.98 (d, J=3.4 Hz, 2H), 4.44 (t, J=3.3 Hz, 2H),3.81 (t, J=8.4 Hz, 4H), 3.74 (t, J=8.1 Hz, 2H), 3.57 (dd, J=8.1, 3.1 Hz,2H), 3.50-3.41 (m, 2H). LC-MS: m/z=403.2 (M+H⁺).

Example 2. Preparation of Compounds 2-4

Compounds 2-4 are prepared as described in Method A using theappropriate bis-boronic acids using a similar procedure as described inExample 1.

LC-MS: m/z Compound IUPAC name ¹H-NMR (M + H⁺) 2 (2R,3S,4R,5S,6R)-2-(400 MHz, CD₃OD) δ 7.59 (d, J = 17.3 Hz, 403.3 (Hydroxymethyl)-6-[3-1H), 7.46-7.38 (m, [(2R,3S,4R,5S,6R)-3,4,5- 3H), 4.99 (d, J = 3.5 Hz,2H), trihydroxy-6- 4.47 (t, J = 3.3 Hz, 2H),(hydroxymethyl)tetrahydropyran- 3.86-3.82 (m, 4H), 3.74 (t, J = 8.1 Hz,2- 2H), 3.59 (dd, J = 8.1, 3.1 Hz, yl]phenyl]tetrahydropyran- 2H), 3.46(ddd, J = 8.2, 5.7, 3,4,5-triol 3.9 Hz, 2H). 3 (2R,3S,4R,5S,6R)-2- (400MHz, CD₃OD) δ 7.86 (s, 471.3 (Hydroxymethyl)-6-[3- 1H), 7.76 (s, 2H),4.98 (d, J = 5.2 Hz, (trifluoromethyl)-5- 2H), 4.30 (dd, J = 5.2,[(2R,3S,4R,5S,6R)-3,4,5- 3.1 Hz, 2H), 3.94 (dd, J = 12.0, trihydroxy-6-7.4 Hz, 2H), 3.84-3.76 (m, (hydroxymethyl)tetrahydropyran- 4H), 3.66(dd, J = 6.9, 3.1 Hz, 2- 2H), 3.57 (td, J = 7.1, 3.1 Hz,yl]phenyl]tetrahydropyran- 2H). 3,4,5-triol 4 (2R,3S,4R,5S,6R)-2- (400MHz, CDCl₃) δ 7.61 (d, J = 1.9 Hz, 461.4 (Hydroxymethyl)-6-[4- 1H), 7.37(dd, J = 8.6, isopropoxy-3- 1.8 Hz, 1H), 7.02 (d, J = 8.7 Hz,[(2R,3S,4R,5S,6R)-3,4,5- 1H), 5.22 (d, J = 5.5 Hz, 1H), trihydroxy-6-4.94 (d, J = 3.5 Hz, 1H), (hydroxymethyl)tetrahydropyran- 4.71-4.61 (m,1H), 4.49 (dd, J = 5.5, 2- 3.1 Hz, 1H), 4.44 (t, J = 3.3 Hz,yl]phenyl]tetrahydropyran- 1H), 3.95 (dd, J = 11.7, 6.6 Hz, 3,4,5-triol1H), 3.89-3.69 (m, 7H), 3.64 (dd, J = 8.1, 3.0 Hz, 1H), 3.53-3.42 (m,1H), 1.36 (dd, J = 10.9, 6.0 Hz, 6H).

Example 3: Preparation of Compound 5 (Method A)(2R,2′R,3S,3′S,4R,4′R,5S,5′S,6R,6′R)-6,6′-([1,1′-Biphenyl]-3,4′-diyl)bis(2-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol)

Step I:((2R,3S,4R,5S,6R)-3-Acetoxy-4,5-dihydroxy-6-(3′-((2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)-[1,1′-biphenyl]-4-yl)tetrahydro-2H-pyran-2-yl)methylAcetate

A mixture of Intermediate B (30 mg, 0.066 mmol),(2R,3S,4R,5S,6R)-2-(3-bromophenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol(C, 30.71 mg, 0.087 mmol), PdCl₂(dppf). CH₂Cl₂ (4.1 mg, 0.005 mmol) andpotassium phosphate (35.7 mg, 0.167 mmol) in degassed DMF is heated in a4 mL sealed vial at 90° C. for 4h. The mixture is filtered through a padof celite and purified directly by reverse phase HPLC on Phenomenex C18Gemini AXIA 5μ, 110A 21.2×75 mm 0% ACN/H₂O+0.01% TFA-To 50% ACN+0.01%TFA in 20 min-To 100% ACN in 5 min-Hold 5 min at 100% ACN. Total RunTime: 30 min to afford the title product.

Step II: Compound 5

((2R,3S,4R,5S,6R)-3-acetoxy-4,5-dihydroxy-6-(3′-((2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)-[1,1′-biphenyl]-4-yl)tetrahydro-2H-pyran-2-yl)methylacetate from Step I is dissolved in MeOH (0.5 mL) and MeONa in MeOH (5μL of 25% w/v, 0.023 mmol) is added and the reaction mixture is stirredat RT for 15h. The reaction is neutralized by the addition of AmberliteIR120H resin until the pH changed to neutral. The reaction mixture isfiltered and the filtrate is evaporated to afford the title compound asa white solid (10.5 mg). ¹H NMR (400 MHz, CD₃OD) δ 7.78 (s, 1H), 7.66(d, J=8.4 Hz, 2H), 7.55 (d, J=8.0 Hz, 3H), 7.44 (d, J=4.9 Hz, 2H), 5.02(2d, J=3.5 Hz, 2H), 4.47 (dd, J=6.2, 3.1 Hz, 2H), 3.88-3.79 (m, 4H),3.74 (td, J=8.0, 2.8 Hz, 2H), 3.61 (td, J=7.9, 3.1 Hz, 2H), 3.56-3.43(m, 2H). LC-MS: m/z=479.4 (M+H⁺)

Example 4. Preparation of Compound 6 (Method A)(2R,2′R,3S,3′S,4R,4′R,5S,5′S,6R,6′R)-6,6′-([1,1′-Biphenyl]-4,4′-diyl)bis(2-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol)

The title compound is prepared using a similar procedure as describedfor Example 3, Compound 5 but using Intermediate A and Intermediate B asstarting materials. ¹H NMR (400 MHz, CD₃OD) δ 7.64 (t, J=10.1 Hz, 4H),7.56 (d, J=8.2 Hz, 4H), 5.03 (d, J=3.3 Hz, 2H), 4.48 (t, J=3.3 Hz, 2H),3.89-3.81 (m, 4H), 3.76 (t, J=8.1 Hz, 2H), 3.62 (dd, J=8.1, 3.0 Hz, 2H),3.56-3.46 (m, 2H). LC-MS: m/z=479.5 (M+H⁺).

Alternative Synthesis of Compound 6 (Method B)

(2R,2′R,3S,3′S,4R,4′R,5S,5′S,6R,6′R)-6,6′-([1,1′-Biphenyl]-4,4′-diyl)bis(2-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol)

Step I:[(2R,3R,4R,5R,6R)-3,4,5-Tris(2,2-dimethylpropanoyloxy)-6-[4-[4-[(2R,3R,4R,5R,6R)-3,4,5-tris(2,2-dimethylpropanoyloxy)-6-(2,2-dimethylpropanoyloxymethyl)tetrahydropyran-2-yl]phenyl]phenyl]tetrahydropyran-2-yl]methyl2,2-dimethylpropanoate

A solution of n-Bu₃MgLi (2.195 mL of 0.66 M, 1.449 mmol) inhexane-heptane-dibutylether (8:20:3) is added to heterogeneous mixtureof 1-iodo-4-(4-iodophenyl)benzene (701 mg, 1.726 mmol) in toluene (4.0mL) and dibutylether (2.4 mL) at 0° C. and stirred at the sametemperature for 3.5 h. A solution of ZnBr₂—LiBr in dibutyl ether (2.17mL of 1.05 M, 2.28 mmol) is added dropwise, cooling bath removed andstirred at RT for 1 h. A solution of[(2R,3R,4S,5S,6R)-6-bromo-3,4,5-tris(2,2-dimethylpropanoyloxy)tetrahydropyran-2-yl]methyl2,2-dimethylpropanoate (2 g, 3.451 mmol, Ref. Sebastien Lemaire et. al.Org. Letts. 2012, 14, 1480-1483) in toluene (3.6 mL) is added, it isplaced on pre-heated oil bath at 100° C. and stirred for 30 h. Thereaction mixture is cooled to RT and poured into aqueous 1N HCl solution(40 mL), extracted with EtOAc (3×25 mL), combined extracts are washedwith brine, dried (Na₂SO₄) and concentrated. The residue is purified onBiotage™ SNAP 100 g silica gel cartridge using EtOAc in Hex (0% to 15%,8 CV) as eluent to afford the title compound (400 mg, 0.347 mmol, 20%)as light yellow solid. ¹H NMR (400 MHz, CDCl₃) δ 7.67 (d, J=8.5 Hz, 4H),7.63 (d, J=8.4 Hz, 4H), 6.07 (t, J=2.8 Hz, 2H), 5.54 (t, J=9.2 Hz, 2H),5.19 (dd, J=9.4, 2.9 Hz, 2H), 5.13 (d, J=2.6 Hz, 2H), 4.32 (dd, J=12.2,5.6 Hz, 2H), 4.19 (dd, J=12.2, 1.9 Hz, 2H), 3.87-3.79 (m, 2H), 1.28 (s,36H), 1.19 (s, 18H), 1.15 (s, 18H).

Step II, Compound 6

To a stirred suspension of[(2R,3R,4R,5R,6R)-3,4,5-tris(2,2-dimethylpropanoyloxy)-6-[4-[4-[(2R,3R,4R,5R,6R)-3,4,5-tris(2,2-dimethylpropanoyloxy)-6-(2,2-dimethylpropanoyloxymethyl)tetrahydropyran-2-yl]phenyl]phenyl]tetrahydropyran-2-yl]methyl2,2-dimethylpropanoate (165 mg, 0.1433 mmol) in MeOH (1.8 mL) is addedMeONa in MeOH (57.0 μL of 0.5 M, 0.287 mmol), stirred at RT for 66 h andquenched with acetic acid (50 μL, 0.8792 mmol). The reaction mixture isconcentrated and directly loaded onto 3 g C18 samplet using DMSO-CH₂Cl₂,dried under house vacuum, purified on Biotage™ SNAP 25 g C18 silica gelcartridge using CH₃CN in water (0% to 50%, 12 CV) as eluent to affordtitle compound (30 mg, 0.0613 mmol, 43%) as white solid. ¹H NMR (400MHz, CD₃OD) δ 7.64 (d, J=8.4 Hz, 4H), 7.55 (d, J=8.2 Hz, 4H), 5.01 (d,J=3.5 Hz, 2H), 4.46 (t, J=3.3 Hz, 2H), 3.83 (d, J=4.7 Hz, 4H), 3.74 (t,J=8.1 Hz, 2H), 3.60 (dd, J=8.1, 3.1 Hz, 2H), 3.53-3.46 (m, 2H)

Example 5. Preparation of Compound 7(3-[(2R,3S,4R,5S,6R)-3,4,5-Trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]-N-[4-[[3-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]benzoyl]amino]phenyl]benzamide)

To a solution of Intermediate D (40 mg, 0.14 mmol), benzene-1,4-diamine(7.6 mg, 0.07 mmol) and HATU (69.20 mg, 0.18 mmol) in DMF (350 μL) isadded triethylamine (29 μL, 0.21 mmol) and the reaction mixture isstirred at RT for 18 h. The mixture is purified directly by reversephase HPLC on Phenomenex C18 Gemini AXIA Pack 5μ 110A 21.2×250 mm (Hold10 min at 10%) 10% ACN/H₂O+0.01% TFA-To 60% ACN+0.01% TFA in 20 min-To100% ACN in 5 min-Hold 5 min at 100% ACN. Total Run Time: 40 min toafford the title product (27.1 mg, 0.04 mmol, 57.22%) as a white solid.¹H NMR (400 MHz, CD₃OD) δ 8.06 (s, 2H), 7.86 (d, J=7.8 Hz, 2H), 7.72 (d,J=7.0 Hz, 6H), 7.53 (t, J=7.8 Hz, 2H), 5.02 (d, J=4.4 Hz, 2H), 4.46-4.39(m, 2H), 3.97-3.75 (m, 6H), 3.68 (dd, J=7.4, 3.0 Hz, 2H), 3.57 (td,J=6.9, 3.0 Hz, 2H). LC-MS: m/z=641.4 (M+H⁺).

Example 6. Preparation of Compounds 8-16

Compounds 8-16 are prepared starting from the appropriate diamine andIntermediate D using a similar procedure as described in Example 5.

LC-MS Compound m/z No. Structure and IUPAC name (M + H⁺) 8(R,R,S,R,S)-N,N′-(Ethane-1,2-diyl)bis(3- 593.4((2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2- yl)benzamide) 9(R,R,S,R,S)-N,N′-(Propane-1,3-diyl)bis(3- 607.4((2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2- yl)benzamide) 10(R,R,S,R,S)-N,N′-(Butane-1,4-diyl)bis(3- 621.4((2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2- yl)benzamide) 11(R,R,S,R,S)-N,N′-(Pentane-1,5-diyl)bis(3- 635.5((2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2- yl)benzamide) 12(R,R,S,R,S)-N,N′-(Hexane-1,6-diyl)bis(3- 649.6((2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2- yl)benzamide) 13(R,R,S,R,S)-N,N′-(Heptane-1,7-diyl)bis(3- 663.5((2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2- yl)benzamide) 14(R,R,S,R,S)-N,N′-(Octane-1,8-diyl)bis(3- 667.5((2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2- yl)benzamide) 15(R,R,S,R,S)-N,N′-((Ethane-1,2- 681.5diylbis(oxy))bis(ethane-2,1-diyl))bis(3-((2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2- yl)benzamide) 16((R,R,S,R,S)-N,N′-(Methylenebis(4,1- 731.5phenylene))bis(3-((2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro- 2H-pyran-2-yl)benzamide)

Example 7. Preparation of Compound 17((2R,3S,4R,5S,6R)-2-(Hydroxymethyl)-6-[3-[(2R,3S,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]oxyphenyl]tetrahydropyran-3,4,5-triol)

Step I:[(2R,3R,4R,5R,6R)-3,4,5-Triacetoxy-6-[3-[(2R,3S,4S,5R,6R)-3,4,5-triacetoxy-6-(acetoxymethyl)tetrahydropyran-2-yl]oxyphenyl]tetrahydropyran-2-yl]methylacetate

To a cold (0° C.) stirred solution of(2R,3R,4R,5R,6R)-2-(acetoxymethyl)-6-(3-hydroxyphenyl)tetrahydro-2H-pyran-3,4,5-triyltriacetate(100 mg, 0.2356 mmol) and[(2R,3R,4S,5S,6R)-3,4,5,6-tetraacetoxytetrahydropyran-2-yl]methylacetate (92 mg, 0.2356 mmol) in CH₂Cl₂ (1.379 mL) is added neat BF₃.OEt₂(100 mg, 0.7068 mmol). The cooling bath is removed, stirred at RT for 15min and then it is heated at 40° C. for 18 h. It is cooled to RT andpoured into aqueous NaHCO₃ solution (4 mL), stirred for 30 min, aqueoussolution is extracted with methylene chloride (2×4 mL). The combinedextracts are dried and concentrated, and purified on Biotage™ SNAP 25 gsilica gel cartridge using EtOAc in Hex (30% to 70%, 8 CV, 70% 6 CV) aseluent to afford the title compound (130 mg, 0.1723 mmol, 73.12%) ashalf-white solid. ¹H NMR (400 MHz, CDCl₃) δ 7.41-7.33 (m, 1H), 7.23 (s,1H), 7.19 (d, J=7.7 Hz, 1H), 7.10 (dd, J=8.2, 2.4 Hz, 1H), 5.92 (t,J=3.2 Hz, 1H), 5.59-5.52 (m, 2H), 5.45 (dd, J=3.5, 1.8 Hz, 1H), 5.39 (t,J=10.1 Hz, 1H), 5.32 (t, J=8.7 Hz, 1H), 5.13 (dd, J=8.9, 3.1 Hz, 1H),5.07 (d, J=3.1 Hz, 1H), 4.43-4.35 (m, 1H), 4.34-4.26 (m, 1H), 4.21-4.04(m, 3H), 3.88-3.75 (m, 1H), 2.21 (s, 3H), 2.165 (s, 3H), 2.153 (s, 3H),2.064 (s, 3H), 2.06 (s, 3H), 2.049 (s, 3H), 2.04 (s, 3H), 2.037 (s, 3H)

Step II: Compound 17

To a stirred solution of[(2R,3R,4R,5R,6R)-3,4,5-triacetoxy-6-[3-[(2R,3S,4S,5R,6R)-3,4,5-triacetoxy-6-(acetoxymethyl)tetrahydropyran-2-yl]oxyphenyl]tetrahydropyran-2-yl]methylacetate (118 mg, 0.1564 mmol) in MeOH (7 mL) is added MeONa (313 μL of0.5 M, 0.1564 mmol) and the reaction mixture is stirred at RT overnight.Dowex 50 WX4-400 ion-exchange resin (H+)[washed several times with MeOHuntil colorless, dried] is added until pH 5-6, filtered off the resinand filtrate is concentrated. The residue is dissolved in CH₃CN-water,freeze dried to the title compound (55 mg, 79%) as white solid. ¹H NMR(400 MHz, CD₃OD) δ 7.21 (t, J=8.0 Hz, 1H), 7.17 (s, 1H), 7.03 (d, J=7.2Hz, 1H), 6.94 (dd, J=8.2, 2.4 Hz, 1H), 5.40 (d, J=1.7 Hz, 1H), 4.85 (d,J=3.3 Hz, 1H), 4.32 (t, J=3.3 Hz, 1H), 3.90 (dd, J=3.4, 1.8 Hz, 1H),3.85-3.44 (m, 9H), 3.43-3.30 (m, 1H). LC-MS: m/z=491.2 (M+H⁺).

Example 8. Preparation of Compound 18((2R,3S,4S,5S,6S)-2-(Hydroxymethyl)-6-[4-[4-[1-[(2S,3S,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]triazol-4-yl]phenyl]triazol-1-yl]tetrahydropyran-3,4,5-triol)

Step I:[(2R,3R,4S,5S,6S)-3,4,5-Triacetoxy-6-[4-[4-[1-[(2S,3S,4S,5R,6R)-3,4,5-triacetoxy-6-(acetoxymethyl)tetrahydropyran-2-yl]triazol-4-yl]phenyl]triazol-1-yl]tetrahydropyran-2-yl]methylacetate

To a stirred solution of[(2R,3R,4S,5S,6S)-3,4,5-triacetoxy-6-azido-tetrahydropyran-2-yl]methylacetate (50 mg, 0.1339 mmol) in DMF (5 mL) are added, copper iodide(5.10 mg, 0.02678 mmol) and DIPEA (25.95 mg, 35 μL, 0.201 mmol) and itis purged with nitrogen gas. The reaction mixture is stirred at 80° C.for 5 h. It is cooled to RT, poured into water, extracted with EtOAc,combined extracts are washed with brine, dried (Na₂SO₄), concentrated.The residue is purified on Biotage™ SNAP 12 g silica gel cartridge usingEtOAc in Hex (15% to 80%) as eluent to afford the title compound (20 mg,34.2%) as light yellow solid. LC-MS: m/z=873.57 (M+H⁺)

Step II: Compound 18

To a stirred suspension of[(2R,3R,4S,5S,6S)-3,4,5-triacetoxy-6-[4-[4-[1-[(2S,3S,4S,5R,6R)-3,4,5-triacetoxy-6-(acetoxymethyl)tetrahydropyran-2-yl]triazol-4-yl]phenyl]triazol-1-yl]tetrahydropyran-2-yl]methylacetate (20 mg, 0.02292 mmol) in MeOH (1.000 mL) is added MeONa (45.84μL of 0.5 M, 0.02292 mmol), suspension became clear and the productstarted crashing out. The reaction mixture is stirred at RT overnight,acetic acid is added, concentrated, suspended in 0.5 mL MeOH, andfiltered to give an off white solid. The precipitate is washed with MeOH(0.5 mL), dried to the title compound (10.5 mg, 79.4%) as white solid.¹H NMR (400 MHz, DMSO-D₆) δ 8.66 (s, 2H), 7.95 (s, 4H), 6.04 (s, 2H),5.32 (d, J=5.0 Hz, 2H), 5.08-4.96 (m, 4H), 4.58 (t, J=5.9 Hz, 2H),3.95-3.86 (m, 2H), 3.75 (dd, J=10.9, 5.5 Hz, 2H), 3.65-3.56 (m, 2H),3.55-3.36 (m, 6H). LC-MS: m/z=537.42 (M+H⁺).

Example 9. Preparation of Compound 19((2R,3S,4S,5S,6S)-2-(Hydroxymethyl)-6-[4-[3-[1-[(2S,3S,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]triazol-4-yl]phenyl]triazol-1-yl]tetrahydropyran-3,4,5-triol)

Step I:[(2R,3R,4S,5S,6S)-3,4,5-Triacetoxy-6-[4-[3-[1-[(2S,3S,4S,5R,6R)-3,4,5-triacetoxy-6-(acetoxymethyl)tetrahydropyran-2-yl]triazol-4-yl]phenyl]triazol-1-yl]tetrahydropyran-2-yl]methylAcetate

To a stirred solution of[(2R,3R,4S,5S,6S)-4,5-diacetoxy-2-(acetoxymethyl)-6-azido-tetrahydropyran-3-yl]acetate(96.6 mg, 0.259 mmol) and 1,3-diethynylbenzene (11 mg, 0.087 mmol) inEtOH (1.1 mL) and water (275 μL) is added sequentially CuSO₄ (5.6 mg,0.035 mmol) and sodium ascorbate (12.3 mg, 0.07 mmol) at RT. Thereaction mixture is stirred at RT over weekend, it is diluted withmethylene chloride, filtered through phase separator, and aqueoussolution is extracted with methylene chloride. The combined extracts areconcentrated, purified on Biotage™ SNAP 12 g silica gel cartridge usingEtOAc in Hex (35% to 75%, 15 CV; 75%, 9 CV) as eluent to afford thetitle compound (69 mg, 90.7%) as white solid. ¹H NMR (400 MHz, CDCl₃) δ8.34 (t, J=1.5 Hz, 1H), 8.07 (s, 2H), 7.89 (dd, J=7.8, 1.7 Hz, 2H), 7.55(t, J=7.8 Hz, 1H), 6.10 (d, J=2.9 Hz, 2H), 6.06-6.02 (m, 2H), 5.97 (dd,J=8.8, 3.7 Hz, 2H), 5.40 (t, J=8.8 Hz, 2H), 4.42 (dd, J=12.5, 5.5 Hz,2H), 4.13-4.05 (m, 2H), 3.99-3.92 (m, 2H), 2.20 (s, 6H), 2.11 (s, 6H),2.09 (s, 6H), 2.08 (s, 6H).

Step II: Compound 19

To a stirred solution of[(2R,3R,4S,5S,6S)-3,4,5-triacetoxy-6-[4-[3-[1-[(2S,3S,4S,5R,6R)-3,4,5-triacetoxy-6-(acetoxymethyl)tetrahydropyran-2-yl]triazol-4-yl]phenyl]triazol-1-yl]tetrahydropyran-2-yl]methylacetate (68 mg, 0.07791 mmol) in MeOH (3.4 mL) is added MeONa (156 μL of0.5 M, 0.078 mmol) in MeOH. A white precipitate is started formingwithin an h. Reaction mixture is stirred at RT overnight. It is filteredoff the fine white precipitate, washed with dry MeOH (2 mL), dried invacuum to afford the title compound as white solid. ¹H NMR (400 MHz,DMSO-D₆) δ 8.80 (s, 2H), 8.41 (s, 1H), 7.85 (dd, J=7.7, 1.6 Hz, 2H),7.53 (t, J=7.8 Hz, 1H), 5.93 (d, J=4.5 Hz, 2H), 5.31 (d, J=5.4 Hz, 2H),5.10 (d, J=5.2 Hz, 2H), 5.02 (d, J=5.4 Hz, 2H), 4.61 (t, J=5.8 Hz, 2H),4.46 (dd, J=8.3, 4.8 Hz, 2H), 3.85 (dd, J=8.5, 6.5 Hz, 2H), 3.70-3.51(m, 6H), 3.40 (td, J=6.8, 3.0 Hz, 2H).

¹H NMR (400 MHz, DMSO-D₆+D₂O) δ 8.68 (s, 2H), 8.33 (s, 1H), 7.82 (dd,J=7.7, 1.5 Hz, 2H), 7.53 (t, J=7.8 Hz, 1H), 5.93 (d, J=4.1 Hz, 2H), 4.46(t, J=3.7 Hz, 2H), 3.84 (dd, J=7.2, 3.3 Hz, 2H), 3.65-3.51 (m, 6H),3.37-3.27 (m, 2H). LC-MS: m/z=537.45 (M+H⁺).

Example 10. Preparation of Compound 20((2R,2′R,3S,3′S,4S,4′S,5S,5′S,6S,6′S)-6,6′-(4,4′-((Prop-2-yn-1-ylazanediyl)bis(methylene))bis(1H-1,2,3-triazole-4,1-diyl))bis(2-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol)

To a solution of[(2R,3R,4S,5S,6S)-4,5-diacetoxy-2-(acetoxymethyl)-6-azido-tetrahydropyran-3-yl]acetate(96.6 mg, 0.259 mmol) and N,N-bis(prop-2-ynyl)prop-2-yn-1-amine (10.3mg, 11 μL, 0.0784 mmol) in EtOH (966 μL) and water (242 μL) issequentially added CuSO₄ (5.0 mg, 0.031 mmol) and sodium ascorbate (11.1mg, 0.0627 mmol). The reaction mixture is stirred at RT for 20 h andevaporated to dryness. The residue is purified by flash columnchromatography on silica gel (2 to 20% MeOH in CH₂Cl₂) to give 73 mg ofa mixture of the desired product that is used as is for the next step.

To a solution of the material obtained in the previous step in MeOH (2mL) is added and MeONa (29 μL of 0.5 M, 0.0146 mmol) in MeOH. Themixture is stirred at RT overnight, AcOH (1 μL, 0.015 mmol) is added andthe mixture is evaporated to dryness. The residue is purified by reversephase HPLC to give 7 mg of the title compound. ¹H NMR (400 MHz, CD₃OD) δ8.23 (s, 2H), 5.96 (s, 2H), 4.60 (s, 2H), 4.30 (s, 4H), 3.95 (dd, J=8.1,3.2 Hz, 2H), 3.82 (s, 2H), 3.69 (m, 6H), 3.25 (m, 2H), 3.13 (s, 1H).

Example 11. Preparation of Compound 21((2R,3S,4R,5S,6R)-2-(Hydroxymethyl)-6-(4-(1-((2S,3S,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)-1H-1,2,3-triazol-4-yl)phenyl)tetrahydro-2H-pyran-3,4,5-triol)

This compound is prepared from Intermediate F and[(2R,3R,4S,5S,6S)-4,5-diacetoxy-2-(acetoxymethyl)-6-azido-tetrahydropyran-3-yl]acetateusing a similar procedure as described in Example 10. ¹H NMR (400 MHz,DMSO-D₆) δ 8.66 (s, 1H), 7.80 (d, J=8.3 Hz, 2H), 7.44 (d, J=8.3 Hz, 2H),5.88 (d, J=4.4 Hz, 1H), 4.68 (d, J=5.3 Hz, 1H), 4.41 (m, 1H), 4.03 (dd,J=5.3, 3.1 Hz, 1H), 3.81 (dd, J=6.7, 3.2 Hz, 1H), 3.66-3.46 (m, 6H),3.46-3.28 (m, 3H). LC-MS: m/z=470.36 (M+H⁺)

Example 12. Preparation of Compound 22 and Compound 23 (Modified MethodD)((2R,3S,4R,5S,6R)-2-(Hydroxymethyl)-6-[2-[4-[2-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]ethyl]phenyl]ethyl]tetrahydropyran-3,4,5-triol)and(2R,3S,4R,5S,6R)-2-(Hydroxymethyl)-6-[4-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]butyl]tetrahydropyran-3,4,5-triol)

Step I

To a solution of Intermediate G (104 mg, 0.1895 mmol) in THF (4 mL) areadded 1,4-diiodobenzene (27.8 mg, 0.0842 mmol), PdCl₂(dppf)-CH₂Cl₂ (3.5mg, 0.0043 mmol), CuI (3.2 mg, 0.0168 mmol) and DIPEA (374, 0.2106mmol). The mixture is stirred at 50° C. overnight under nitrogen. Afterremoval of the solvent under reduced pressure, the residue is separatedon Biotage™ SNAP 25 g silica gel cartridge using a gradient of EtOAc inHex 0-15% in 20 column volume to obtain an inseparable ˜2:1 mixture (92mg) of(2R,3R,4R,5R,6R)-3,4,5-tribenzyloxy-2-(benzyloxymethyl)-6-[2-[4-[2-[(2R,3R,4R,5R,6R)-3,4,5-tribenzyloxy-6-(benzyloxymethyl)tetrahydropyran-2-yl]ethynyl]phenyl]ethynyl]tetrahydropyranand(2R,3R,4R,5R,6R)-3,4,5-tribenzyloxy-2-(benzyloxymethyl)-6-[4-[(2R,3R,4R,5R,6R)-3,4,5-tribenzyloxy-6-(benzyloxymethyl)tetrahydropyran-2-yl]buta-1,3-diynyl]tetrahydropyran,respectively, which is used directly in the next step without furtherpurification. LC-MS: m/z=1193.9 (M+Na⁺). LC-MS: m/z=1118.9 (M+Na⁺).

Step II: Compound 22 and Compound 23

To a solution of the ˜2:1 mixture from Step I (92 mg) in MeOH (5 mL) areadded a catalytic amount of 20% Pd(OH)₂/C and a drop of acetic acid. Themixture is hydrogenated using a hydrogen balloon and stirred at RTovernight. After filtration, the solvent is removed and the residue ispurified using reverse phase HPLC to obtain Compound 22 (11 mg) andCompound 23 (9 mg), both as white solid. Compound 22: ¹H NMR (400 MHz,CD₃OD) δ 7.13 (s, 4H), 3.81 (m, 4H), 3.75-3.53 (m, 8H), 3.45 (m, 2H),2.75 (m, 2H), 2.66-2.50 (m, 2H), 2.02 (m, 2H), 1.70 (m, 2H). LC-MS:m/z=459.4 (M+H⁺). Compound 23: ¹H NMR (400 MHz, CD₃OD) δ 3.83 (d, 2H),3.77 (m, 2H), 3.72-3.61 (m, 6H), 3.58 (m, 2H), 3.39 (m, 2H), 1.76 (m,2H), 1.45 (m, 6H). LC-MS: m/z=383.3 (M+H⁺).

Example 13. Preparation of Compound 24((2R,3S,4R,5S,6R)-2-(Hydroxymethyl)-6-[(E)-3-[4-[(E)-3-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]prop-1-enyl]phenyl]allyl]tetrahydropyran-3,4,5-triol)

Step I:[(2R,3R,4R,5R,6R)-3,4,5-Triacetoxy-6-[(E)-3-[4-[(E)-3-[(2R,3R,4R,5R,6R)-3,4,5-triacetoxy-6-(acetoxymethyl)tetrahydropyran-2-yl]prop-1-enyl]phenyl]allyl]tetrahydropyran-2-yl]methylAcetate

To a solution of Intermediate H (112.9 mg, 0.303 mmol) in DMF (1.5 mL)are added 1,4-diiodobenzene (50 mg, 0.152 mmol), palladium acetate (3.4mg, 0.0152 mmol), tetrabutylammonium bromide (48.9 mg, 0.152 mmol) andsodium bicarbonate (38.2 mg, 0.455 mmol). The reaction mixture is heatedat 85° C. overnight under N₂, concentrated and purified on Biotage™ SNAP(10 g silica gel cartridge) using EtOAc in Hex 0 to 70% as eluent in 20column volume to afford title compound (71 mg, 57% yield). LC-MS:m/z=819.5 (M+H⁺).

In addition,(2R,3R,4R,5R,6R)-2-(acetoxymethyl)-6-((E)-3-(4-iodophenyl)allyl)tetrahydro-2H-pyran-3,4,5-triyltriacetate (33 mg, 20% yield) is isolated as a minor compound. LC-MS:m/z=575.3 (M+H⁺).

Step II: Compound 24

To a stirred solution of[(2R,3R,4R,5R,6R)-3,4,5-triacetoxy-6-[(E)-3-[4-[(E)-3-[(2R,3R,4R,5R,6R)-3,4,5-triacetoxy-6-(acetoxymethyl)tetrahydropyran-2-yl]prop-1-enyl]phenyl]allyl]tetrahydropyran-2-yl]methylacetate (70 mg, 0.0855 mmol) in MeOH (1.1 mL) is added 2 drops of a 25%wt/v MeONa in MeOH. The reaction mixture is stirred at RT for 3h. Themixture is diluted with MeOH and neutralized with resin Amberlite 120(H). After filtration the resin is washed with MeOH, MeOH/H₂O (2:1, v/v)then a mixture dioxane/MeOH (1:1, v/v). The beige residue is trituratedwith MeOH to afford 19 mg of expected compound which is repurified bypreparative HPLC to afford(2R,3S,4R,5S,6R)-2-(hydroxymethyl)-6-[(E)-3-[4-[(E)-3-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]prop-1-enyl]phenyl]allyl]tetrahydropyran-3,4,5-triol(12.9 mg, 31% yield). ¹H NMR (400 MHz, CD₃OD) δ 7.21 (s, 4H), 6.38 (d,2H), 6.28-6.10 (m, 2H), 3.88 (dd, 2H), 3.76-3.59 (m, 8H), 3.55 (t, 2H),3.43 (ddd, 2H), 2.62-2.46 (m, 2H), 2.48-2.30 (m, 2H). LC-MS: m/z=483.4(M+H⁺)

Example 14. Preparation of Compound 25((2R,3S,4R,5S,6R)-2-(Hydroxymethyl)-6-[(E)-3-[3-[(E)-3-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]prop-1-enyl]phenyl]allyl]tetrahydropyran-3,4,5-triol)

Compound 25 is prepared according to a similar procedure as described inExample 13, Compound 24 using 1,3-diiodobenzene instead of1,4-diiodobenzene as starting material. ¹H NMR (400 MHz, CD₃OD) δ 7.30(s, 1H), 7.12 (d, 3H), 6.40 (d, 2H), 6.30-6.10 (m, 2H), 3.90 (s, 2H),3.79-3.33 (m, 12H), 2.55 (d, 2H), 2.42 (dd, 2H). LC-MS: m/z=483.4(M+H⁺).

Example 15. Preparation of Compounds 26-33

Compounds 26-33 are prepared from Intermediate H and appropriate1,4-diiodobenzene derivatives using a similar procedure as described inExample 14.

LC-MS m/z Compound IUPAC name ¹H-NMR (M + H⁺) 26 (Methyl 2,5-bis[(E)-3-(400 MHz, CD₃OD) δ 7.70 (d, 541.4 [(2R,3S,4R,5S,6R)-3,4,5- 1H), 7.47(dd, 2H), 7.06 (d, trihydroxy-6- 1H), 6.42 (d, 1H),(hydroxymethyl)tetrahydropyran- 6.35-6.20 (m, 1H), 6.20-6.04 (m, 1H),2-yl]prop-1-enyl]benzoate) 3.91 (d, 2H), 3.79 (s, 3H), 3.75-3.6 (m, 8H),3.55 (td, 2H), 3.50-3.40 (m, 2H), 2.57 (dd, 2H), 2.42 (m, 2H). 27((2R,3S,4R,5S,6R)-2- (400 MHz, CD₃OD) δ 7.37 (d, 497.5(Hydroxymethyl)-6-[(E)-3-[3- 1H), 7.20-7.04 (m, 2H), methyl-4-[(E)-3-6.68 (d, 1H), 6.42 (d, 1H), [(2R,3S,4R,5S,6R)-3,4,5- 6.33-6.19 (m, 1H),6.19-6.06 (m, trihydroxy-6- 1H), 3.97 (m, 2H), 3.80 (m,(hydroxymethyl)tetrahydropyran- 3H), 3.78-3.67 (m, 5H), 2-yl]prop-1-3.63 (td, 2H), 3.57-3.47 (m, 2H), enyl]phenyl]allyl]tetrahydropyran-2.71-2.56 (m, 2H), 2.50 (dt, 3,4,5-triol) 2H), 2.29 (s, 3H 28((2R,3S,4R,5S,6R)-2-[(E)-3-[3- (400 MHz, CD₃OD) δ 7.36 (t, 501.5Fluoro-4-[(E)-3- 1H), 7.02 (dd, 2H), 6.51 (d, [(2R,3S,4R,5S,6R)-3,4,5-1H), 6.37 (d, 1H), trihydroxy-6- 6.33-6.11 (m, 2H), 3.89 (td, 2H),(hydroxymethyl)tetrahydropyran- 3.76-3.59 (m, 8H), 3.55 (td, 2H),2-yl]prop-1-enyl]phenyl]allyl]-6- 3.44 (ddd, 2H), 2.63-2.49 (m,(hydroxymethyl)tetrahydropyran- 2H), 2.49-2.35 (m, 2H). 3,4,5-triol) 29((2R,3S,4R,5S,6R)-2-[(E)-3-[3- (400 MHz, CD₃OD) δ 7.54 (d, 517.4Chloro-4-[(E)-3- 1H), 7.36 (d, 1H), 7.27 (d, 1H),[(2R,3S,4R,5S,6R)-3,4,5- 6.82 (d, 1H), 6.44 (d, 1H), trihydroxy-6- 6.32(dt, 2H), 3.98 (d, 2H), 3.75 (tdt, (hydroxymethyl)tetrahydropyran- 8H),3.63 (q, 2H), 2-yl]prop-1-enyl]phenyl]allyl]-6- 3.58-3.45 (m, 2H),2.74-2.58 (m, 2H), (hydroxymethyl)tetrahydropyran- 2.58-2.41 (m, 2H).3,4,5-triol) 30 ((2R,3S,4R,5S,6R)-2-[(E)-3-[2,5- (400 MHz, DMSO) δ 7.01(s, 543.5 Dimethoxy-4-[(E)-3- 2H), 6.61 (d, 2H), (2R,3S,4R,5S,6R)-3,4,5-6.39-6.21 (m, 2H), 3.82-3.65 (m, 8H), trihydroxy-6- 3.62-3.25 (m, 12H),2.73 (m, (hydroxymethyl)tetrahydropyran- 4H).2-yl]prop-1-enyl]phenyl]allyl]-6- (hydroxymethyl)tetrahydropyran-3,4,5-triol) 31 ((2R,3S,4R,5S,6R)-2-[(E)-3-[3,5- (400 MHz, CD₃OD) δ 7.02(s, 511.5 Dimethyl-4-[(E)-3- 2H), 6.38 (d, 1H), [(2R,3S,4R,5S,6R)-3,4,5-6.30-6.10 (m, 1H), 5.90 (ddd, 1H), trihydroxy-6- 5.32 (dd, 1H), 4.36 (d,1H), 3.95 (dd, (hydroxymethyl)tetrahydropyran- 1H), 3.85-3.35 (m, 14H),2-yl]prop-1-enyl]phenyl]allyl]-6- 2.68-2.52 (m, 1H), 2.54-2.38 (m,(hydroxymethyl)tetrahydropyran- 1H), 2.25 (s, 6H). 3,4,5-triol) 32((2R,3S,4R,5S,6R)-2- (400 MHz, CD₃OD) δ 7.76 (d, 528.5(Hydroxymethyl)-6-[(E)-3-[3- 1H), 7.56 (q, 2H), 6.73 (d, 1H),nitro-4-[(E)-3- 6.54-6.21 (m, 3H), 3.91 (s, [(2R,3S,4R,5S,6R)-3,4,5-2H), 3.77-3.60 (m, 8H), trihydroxy-6- 3.55 (q, 2H), 3.49-3.34 (m, 2H),(hydroxymethyl)tetrahydropyran- 2.59 (dd, 2H), 2.45 (s, 2H).2-yl]prop-1- enyl]phenyl]allyl]tetrahydropyran- 3,4,5-triol) 33(2,5-Bis[(E)-3- (400 MHz, CD₃OD) δ 508.5 [(2R,3S,4R,5S,6R)-3,4,5-7.74-7.35 (m, 3H), 6.70 (d, 1H), trihydroxy-6- 6.59-6.24 (m, 3H), 3.91(d, 2H), (hydroxymethyl)tetrahydropyran- 3.77-3.60 (m, 8H), 3.55 (dt,2-yl]prop-1-enyl]benzonitrile) 2H), 3.45 (dt, 2H), 2.77-2.31 (m, 4H).

Example 16. Preparation of Compound 34((2R,3S,4R,5S,6R)-2-(Hydroxymethyl)-6-[(E)-3-[4-[2-[4-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]phenyl]ethynyl]phenyl]allyl]tetrahydropyran-3,4,5-triol)

Step I:[(2R,3R,4R,5R,6R)-3,4,5-Triacetoxy-6-[(E)-3-[4-[2-[4-[(2R,3R,4R,5R,6R)-3,4,5-triacetoxy-6-(acetoxymethyl)tetrahydropyran-2-yl]phenyl]ethynyl]phenyl]allyl]tetrahydropyran-2-yl]methylAcetate

To a mixture of(2R,3R,4R,5R,6R)-2-(acetoxymethyl)-6-((E)-3-(4-iodophenyl)allyl)tetrahydro-2H-pyran-3,4,5-triyltriacetate(33 mg, 0.057 mmol), Intermediate F (24.9 mg, 0.057 mmol), Pd(dppf) (4.7mg, 0.0057 mmol), CuI (2.2 mg, 0.0115 mmol) in DMF (6604) is added NEt₃(24 μL, 0.173 mmol). The system is flushed with nitrogen and the mixtureis heated to 70° C. overnight under N₂. The RM is diluted with EtOAc andH₂O, filtered on celite. The organic phase is washed with brine, driedover Na₂SO₄, filtered, concentrated and purified on a Biotage™ SNAP (10g silica gel cartridge) using a gradient of EtOAc in Hex (10-80%) in 25column volume to afford title compound (22 mg, 43.5%). LC-MS: m/z=879.6(M+H⁺).

Step II: Compound 34

To the residue from step I (22 mg, 0.025 mmol) in MeOH (660 μL) is added2 drops of 25% w/v MeONa in MeOH. The reaction mixture is stirred at RTfor 2h, neutralized with resin Amberlite 120 (H). After filtration, theresin is washed with MeOH and the filtrate concentrated to dryness. Theresidue is purified by reverse phase HPLC to afford the title compound(2.6 mg, 20%). ¹H NMR (400 MHz, CD₃OD) δ 7.49-7.37 (m, 4H), 7.32 (m,4H), 6.43 (d, 1H), 6.36-6.21 (m, 1H), 5.39 (s, 2H), 4.30 (t, 1H), 3.90(t, 1H), 3.82-3.31 (m, 10H), 2.57 (dd, 1H), 2.44 (dd, 1H). LC-MS:m/z=543.4 (M+H⁺).

Example 17. Preparation of Compound 35 (Method F)((2R,3S,4R,5S,6R)-2-(Hydroxymethyl)-6-[2-[3-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]phenyl]ethynyl]tetrahydropyran-3,4,5-triol)

Step I:[(2R,3R,4R,5R,6R)-3,4,5-Triacetoxy-6-[3-[2-[(2R,3R,4R,5R,6R)-3,4,5-tribenzyloxy-6-(benzyloxymethyl)tetrahydropyran-2-yl]ethynyl]phenyl]tetrahydropyran-2-yl]methylAcetate

To a solution of Intermediate G (160 mg, 0.2916 mmol) in DMF (4 mL) areadded Intermediate I (113.7 mg, 0.2333 mmol), PdCl₂(dppf)-CH₂Cl₂ (9.5mg, 0.0117 mmol), CuI (8.9 mg, 0.0467 mmol) and DIPEA (102 μL, 0.5832mmol). The mixture is stirred at 80° C. overnight under nitrogen. Afterremoval of the solvent under reduced pressure, the residue is separatedon Biotage™ SNAP 25 g silica gel cartridge using a gradient of EtOAc inHex 0-15% in 20 CV to obtain the title compound (65 mg). LC-MS:m/z=955.5 (M+H⁺).

Step II:(2R,3R,4R,5R,6R)-2-(acetoxymethyl)-6-(3-(((2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)ethynyl)phenyl)tetrahydro-2H-pyran-3,4,5-triylTriacetate

To a solution of[(2R,3R,4R,5R,6R)-3,4,5-triacetoxy-6-[3-[2-[(2R,3R,4R,5R,6R)-3,4,5-tribenzyloxy-6-(benzyloxymethyl)tetrahydropyran-2-yl]ethynyl]phenyl]tetrahydropyran-2-yl]methylacetate from Step I (20 mg, 0.0209 mmol) in CH₃CN (2 mL) is added TMSI(41.90 mg, 29.80 μL, 0.2094 mmol) in a sealed vessel. The mixture isstirred at RT overnight. It is then quenched with drops of water. Afterremoval of the solvent under reduced pressure, the residue is useddirectly in the next step without purification.

Step III: Compound 35

To the mixture from step II in MeOH (2 mL) is added a drop of 25% MeONain MeOH. The mixture is stirred at RT for 20 min, then neutralized withresin Amberlite IR 120 (H) and filtered. The filtrate is concentrated todryness and the residue is purified using prep-HPLC to provide(2R,3S,4R,5S,6R)-2-(hydroxymethyl)-6-[2-[3-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]phenyl]ethynyl]tetrahydropyran-3,4,5-triol(1.5 mg) as a white solid. ¹H NMR (400 MHz, CD₃OD) δ 7.51 (s, 1H),7.45-7.35 (m, 1H), 7.28 (m, 2H), 4.82 (d, 1H), 4.76 (d, 1H), 4.24 (m,1H), 3.91 (m, 1H), 3.84 (m, 1H), 3.80-3.75 (m, 2H), 3.74-3.69 (m, 2H),3.67-3.60 (m, 2H), 3.58-3.46 (m, 2H), 3.41 (m, 1H). LC-MS: m/z=427.3(M+H⁺).

Example 18. Preparation of Compound 36((2R,3S,4R,5S,6R)-2-(Hydroxymethyl)-6-[4-[4-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]triazol-1-yl]phenyl]tetrahydropyran-3,4,5-triol)

Step I:[(2R,3S,6S)-3-Acetoxy-6-[4-(tert-butoxycarbonylamino)phenyl]-3,6-dihydro-2H-pyran-2-yl]methylAcetate

To a solution of[(2R,3S,4R)-3,4-diacetoxy-3,4-dihydro-2H-pyran-2-yl]methyl acetate (4 g,14.69 mmol) in CH₃CN (35 mL) are added[4-(tert-butoxycarbonylamino)phenyl]boronic acid (6.965 g, 29.38 mmol)and Pd(OAc)₂ (494.6 mg, 2.203 mmol). The mixture is stirred at RTovernight and then added another batch of[4-(tert-butoxycarbonylamino)phenyl]boronic acid (6.965 g, 29.38 mmol)and Pd(OAc)₂ (495 mg, 2.203 mmol). After stirring at RT overnight, themixture is diluted with CH₂Cl₂ (35 mL), filtered on a pad of celite. Thefiltrate is concentrated to dryness under reduced pressure and theresidue is purified on Biotage™ SNAP 100 g silica gel cartridge using agradient of EtOAc in Hex (0-15%, 20 CV) to obtain the title compound(660 mg) as a glassy solid.

Step II:((2R,3S,6S)-3-acetoxy-6-(4-aminophenyl)-3,6-dihydro-2H-pyran-2-yl)methylAcetate

To a solution of[(2R,3S,6S)-3-acetoxy-6-[4-(tert-butoxycarbonylamino)phenyl]-3,6-dihydro-2H-pyran-2-yl]methylacetate (303 mg, 0.7473 mmol) in 1 mL of CH₂Cl₂ is added TFA (1 mL,12.98 mmol). The mixture is stirred at RT for 20 min. After removal ofthe solvent under reduced pressure, the residue is dissolved in 10 mL ofCH₂Cl₂, washed with 10% sodium bicarbonate (2 mL) and brine (5 mL)consecutively, dried over sodium sulfate, filtered, and concentrated todryness. The residue is used in the next step without furtherpurification. LC-MS of the mixture shows two products with the samedesired mass. LC-MS: m/z=306.4 (M+H⁺).

Step III:[(2R,3S,6S)-3-Acetoxy-6-(4-azidophenyl)-3,6-dihydro-2H-pyran-2-yl]methylAcetate

To a solution of the mixture from Step II in 2 mL of ACN are addedt-BuONO (133 μL, 1.121 mmol) and TMSN₃ (119 μL, 0.8968 mmol) at 0° C.and then stirred at RT for 1h. After removal of the solvent underreduced pressure, the mixture is purified on Biotage™ SNAP 25 g silicagel cartridge using a gradient of EtOAc in Hex 0-15% in 20 CV to obtaina mixture (190 mg), containing title compound as oil, which is useddirectly in the next step without further purification. LC-MS: m/z=356.5(M+Na⁺).

Step IV:((2R,3S,6S)-3-acetoxy-6-(4-(4-((2R,3R,4R,5R,6R)-3,4,5-tris(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-2-yl)-1H-1,2,3-triazol-1-yl)phenyl)-3,6-dihydro-2H-pyran-2-yl)methylAcetate

To a solution of[(2R,3S,6S)-3-acetoxy-6-(4-azidophenyl)-3,6-dihydro-2H-pyran-2-yl]methylacetate (60 mg, 0.1811 mmol) in THF (5 mL) are added(2R,3R,4R,5R,6R)-3,4,5-tribenzyloxy-2-(benzyloxymethyl)-6-ethynyl-tetrahydropyran(99.4 mg, 0.1811 mmol), CuI (6.9 mg, 0.0362 mmol) and DIPEA (24 mg, 32μL, 0.1811 mmol). The mixture is stirred at 45° C. overnight undernitrogen. After removal of the solvent under reduced pressure, theresidue is separated on Biotage™ SNAP 25 g silica gel cartridge using agradient of EtOAc in Hex 0-45% in 20 CV to obtain a mixture (142 mg)containing[(2R,3S,6S)-3-acetoxy-6-[4-[4-[(2R,3R,4R,5R,6R)-3,4,5-tribenzyloxy-6-(benzyloxymethyl)tetrahydropyran-2-yl]triazol-1-yl]phenyl]-3,6-dihydro-2H-pyran-2-yl]methylacetate. LC-MS: m/z=880.7 (M+H⁺).

Step V:((2R,3S,4R,5S,6R)-3-acetoxy-4,5-dihydroxy-6-(4-(4-((2R,3R,4R,5R,6R)-3,4,5-tris(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-2-yl)-1H-1,2,3-triazol-1-yl)phenyl)tetrahydro-2H-pyran-2-yl)methylAcetate

To a solution of[(2R,3S,6S)-3-acetoxy-6-[4-[4-[(2R,3R,4R,5R,6R)-3,4,5-tribenzyloxy-6-(benzyloxymethyl)tetrahydropyran-2-yl]triazol-1-yl]phenyl]-3,6-dihydro-2H-pyran-2-yl]methylacetate (160 mg, 0.1818 mmol) in water (2 mL)/t-BuOH (4 mL) are addedNMO (42.6 mg, 384, 0.3636 mmol), 2.5% OsO₄/t-BuOH (184.9 mg, 973 μL,0.0182 mmol), methanesulfonamide (25.9 mg, 0.2727 mmol) and 2,6-lutidine(19.48 mg, 21 μL, 0.1818 mmol). The mixture is stirred at RT for 5 days.Then it is diluted with CH₂Cl₂ (10 mL), quenched with 10% sodiumbisulfate solution (5 mL), extracted with CH₂Cl₂ (3×10 mL). The combinedorganic extracts are dried over sodium sulfate, filtered, andconcentrated to dryness. The residue is purified on Biotage™ SNAP 25 gsilica gel cartridge using a gradient of MeOH in CH₂Cl₂ 0-5% in 20 CV toobtain a mixture (150 mg) as oil. LC-MS: m/z=914.7 (M+H+).

Step VI: Compound 36

To a solution of the mixture from Step V, containing[(2R,3S,4R,5S,6R)-3-acetoxy-4,5-dihydroxy-6-[4-[4-[(2R,3R,4R,5R,6R)-3,4,5-tribenzyloxy-6-(benzyloxymethyl)tetrahydropyran-2-yl]triazol-1-yl]phenyl]tetrahydropyran-2-yl]methylacetate (60 mg, 0.0657 mmol) in MeOH (2 mL) is added one drop of 25%MeONa/MeOH. The mixture is stirred at RT for 30 min. It is thenneutralized with resin Amberlite IR 120 (H), filtered and concentratedto dryness. The residue is not purified and dissolved in MeOH (2 mL).Then to it are added a catalytic amount of 20% Pd(OH)₂ and a drop ofAcOH. The mixture is hydrogenated using a hydrogen balloon and stirredat RT overnight. After filtration, the solvent is removed under reducedpressure and the residue is purified on reverse phase HPLC to providethe title compound (11 mg) as a white solid. ¹H NMR (400 MHz, CD₃OD) δ8.47 (s, 1H), 7.78 (d, 2H), 7.62 (d, 2H), 5.10 (d, 1H), 4.91 (d, 1H),4.45 (t, 1H), 4.28 (m, 1H), 3.91-3.58 (m, 7H), 3.55 (m, 1H), 3.47 (m,1H), 3.41-3.33 (m, 1H). LC-MS: m/z=470.4 (M+H⁺).

Example 19. Preparation of Compound 37((2R,3S,4R,5S,6R)-2-(Hydroxymethyl)-6-[2-[3-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]phenyl]ethyl]tetrahydropyran-3,4,5-triol)

To a solution of[(2R,3R,4R,5R,6R)-3,4,5-triacetoxy-6-[3-[2-[(2R,3R,4R,5R,6R)-3,4,5-tribenzyloxy-6-(benzyloxymethyl)tetrahydropyran-2-yl]ethynyl]phenyl]tetrahydropyran-2-yl]methylacetate (Example 17, step I) (32 mg, 0.0335 mmol) in MeOH (2 mL) isadded a drop of 25% MeONa/MeOH. The mixture is stirred at RT for 30 min.Then it is neutralized with resin Amberlite IR 120 (H), and filtered.The filtrate is concentrated to dryness and the residue is dissolved inMeOH (2 mL). Then to it are added 20% Pd(OH)₂/C and drop of acetic acid.The mixture is hydrogenated using a hydrogen balloon and stirred at RTovernight. After filtration, it is concentrated to dryness and theresidue is separated using reverse phase prep-HPLC to obtain the titlecompound (11 mg) as a white solid. ¹H NMR (400 MHz, CD₃OD) δ 7.38 (s,1H), 7.27 (d, 2H), 7.21-7.09 (m, 1H), 4.95 (d, 1H), 4.44 (t, 1H),3.94-3.74 (m, 4H), 3.74-3.52 (m, 6H), 3.45 (m, 2H), 2.80 (m, 1H),2.73-2.57 (m, 1H), 2.17-1.95 (m, 1H), 1.82-1.65 (m, 1H). LC-MS:m/z=431.4 (M+H⁺).

Example 20. Preparation of Compound 38(2R,3S,4R,5S,6R)-2-(hydroxymethyl)-6-[3-[4-[3-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]propyl]phenyl]propyl]tetrahydropyran-3,4,5-triol

Step I:[(2R,3R,4R,5R,6R)-3,4,5-Triacetoxy-6-[3-[4-[3-[(2R,3R,4R,5R,6R)-3,4,5-triacetoxy-6-(acetoxymethyl)tetrahydropyran-2-yl]propyl]phenyl]propyl]tetrahydropyran-2-yl]methylAcetate

To[(2R,3R,4R,5R,6R)-3,4,5-triacetoxy-6-[(E)-3-[4-[(E)-3-[(2R,3R,4R,5R,6R)-3,4,5-triacetoxy-6-(acetoxymethyl)tetrahydropyran-2-yl]prop-1-enyl]phenyl]allyl]tetrahydropyran-2-yl]methylacetate (Example 13, Step I) (80 mg, 0.097 mmol) in MeOH (2.4 mL) isadded Pd(OH)₂ (13.7 mg, 0.097 mmol) and 1 drop AcOH. The reactionmixture is hydrogenated under 1 atmosphere of H₂ overnight. The reactionmixture is filtered on celite, washed with MeOH. The filtrate isconcentrated and purified on Biotage™ SNAP 10 g silica gel cartridgeusing EtOAc in Hex (15 to 100%) to provide the title compound (36 mg,44.8%). LC-MS: m/z=823.7 (M+H⁺).

Step II: Compound 38

To a solution of[(2R,3R,4R,5R,6R)-3,4,5-triacetoxy-6-[3-[4-[3-[(2R,3R,4R,5R,6R)-3,4,5-triacetoxy-6-(acetoxymethyl)tetrahydropyran-2-yl]propyl]phenyl]propyl]tetrahydropyran-2-yl]methylacetate (36 mg, 0.097 mmol) in MeOH (1.6 mL) is added catalytic MeONa (2μL of 25% w/v, 0.0097 mmol). The reaction mixture is stirred at RT for1h, neutralized with Amberlite IR120(H). After filtration, the solventis removed under reduced pressure and the residue purified by reversephase HPLC to afford the title compound (6.9 mg, 14%). ¹H NMR (400 MHz,CD₃OD) δ 7.00 (s, 4H), 3.80-3.72 (m, 2H), 3.67 (dd, 2H), 3.63-3.54 (m,4H), 3.55-3.46 (m, 4H), 3.31-3.22 (m, 2H), 2.60-2.40 (m, 4H), 1.64 (ddd,6H), 1.48-1.25 (m, 2H). LC-MS: m/z=487.4 (M+H⁺).

Example 21 Preparation of Compound 39(3-[(2R,3S,4R,5S,6R)-3,4,5-Trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]-N-[4-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]phenyl]benzamide)

Step I:[(2R,3S,6S)-3-Acetoxy-6-[4-[[3-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]benzoyl]amino]phenyl]-3,6-dihydro-2H-pyran-2-yl]methylAcetate

To a mixture of Intermediate D (40.4 mg, 0.142 mmol) and[(2R,3S,6S)-3-acetoxy-6-(4-aminophenyl)-3,6-dihydro-2H-pyran-2-yl]methylacetate (from Step II of Example 18, 43.4 mg, 0.142 mmol) in DMF (2.7mL) is added 2,6-lutidine (49 μL, 0.426 mmol) followed by HATU (59.4 mg,0.156 mmol) at 0° C. The reaction mixture is stirred at RT overnight,diluted with EtOAc, washed with H₂O, brine. The organic phase is driedover Na₂SO₄, filtered and concentrated. The residue is purified byBiotage™ SNAP 10 g silica gel cartridge using MeOH/CH₂Cl₂ (0 to 20%) toafford the title compound (45 mg, 55% yield), which is used in the nextstep without further purification. LC-MS: m/z=572.4 (M+H⁺).

Step II:[(2R,3S,4R,5S,6R)-3-Acetoxy-4,5-dihydroxy-6-[4-[[3-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]benzoyl]amino]phenyl]tetrahydropyran-2-yl]methylAcetate

To a solution of[(2R,3S,6S)-3-acetoxy-6-[4-[[3-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]benzoyl]amino]phenyl]-3,6-dihydro-2H-pyran-2-yl]methylacetate (45 mg, 0.0788 mmol) in a mixture of water (0.45 mL)/t-BuOH(0.45 mL) are added methanesulfonamide (22.5 mg, 0.236 mmol), 2.5%OsO₄/t-BuOH (148.3, 0.012 mmol), NMO (37 mg, 0.315 mmol) and2,6-lutidine (18 μL, 0.157 mmol). The mixture is stirred at RT for 2days, quenched with 15% sodium bisulfite and diluted with EtOAc. Theaqueous phase is separated, the organic phase is washed with water,brine and dried over sodium sulfate. The solvent is removed underreduced pressure to afford the title compound (29 mg, 36% yield). LC-MS:m/z=606.4 (M+H⁺).

Step III: Compound 39

To a solution of[(2R,3S,4R,5S,6R)-3-acetoxy-4,5-dihydroxy-6-[4-[[3-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]benzoyl]amino]phenyl]tetrahydropyran-2-yl]methylacetate (29 mg, 0.048 mmol) in MeOH (0.45 mL) is added 2 drops MeONa 25%w/v in MeOH. The reaction mixture is stirred at RT for 1h, diluted withMeOH, neutralized with resin Amberlite 120 (H), filtered and dried. Theresidue is purified by reverse phase HPLC to provide the title compound(2.7 mg). ¹H NMR (400 MHz, CD₃OD) δ 7.96 (s, 1H), 7.77 (d, 1H), 7.64 (d,2H), 7.58 (d, 1H), 7.49-7.31 (m, 3H), 4.9 (m, 2H), 4.36 (dd, 2H),3.88-3.30 (m, 10H). LC-MS: m/z=522.4 (M+H⁺).

Example 22. Preparation of Compound 40((2R,3S,4R,5S,6R)-2-(Hydroxymethyl)-6-[(E)-3-[3-[2-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]ethynyl]phenyl]allyl]tetrahydropyran-3,4,5-triol)

(2R,3R,4R,5R,6R)-2-(acetoxymethyl)-6-((E)-3-(3-iodophenyl)allyl)tetrahydro-2H-pyran-3,4,5-triyltriacetate is prepared according to the same procedure as described inExample 13 using 1,3-diiodobenzene instead of 1,4-diiodobenzene.

Step I:(2R,3R,4R,5R,6R)-2-(acetoxymethyl)-6-((E)-3-(3-(((2R,3R,4R,5R,6R)-3,4,5-tris(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-2-yl)ethynyl)phenyl)allyl)tetrahydro-2H-pyran-3,4,5-triylTriacetate

To(2R,3R,4R,5R,6R)-2-(acetoxymethyl)-6-((E)-3-(3-iodophenyl)allyl)tetrahydro-2H-pyran-3,4,5-triyltriacetate (53 mg, 0.0923 mmol),trimethyl-[2-[(2R,3R,4R,5R,6R)-3,4,5-tribenzyloxy-6-(benzyloxymethyl)tetrahydropyran-2-yl]ethynyl]silane(Step II of Intermediate G, 57.3 mg, 0.0923 mmol), PdCl₂(dppf)-CH₂Cl₂(7.5 mg, 0.0092 mmol) and CuI (3.5 mg, 0.0185 mmol) in DMF (1.1 mL) areadded H₂O (16.6 μl, 0.923 mmol) and DBU (140.5 mg, 0.923 mmol). Thesystem is flushed with nitrogen and the mixture is heated to 70° C.overnight under N₂. The reaction mixture is diluted with EtOAc and H₂O,filtered on celite. The organic phase is washed with brine, dried overNa₂SO₄, filtered and concentrated to be used in the next step withoutfurther purification.

Step II:(2R,3S,4R,5S,6R)-2-(Hydroxymethyl)-6-[(E)-3-[3-[2-[(2R,3R,4R,5R,6R)-3,4,5-tribenzyloxy-6-(benzyloxymethyl)tetrahydropyran-2-yl]ethynyl]phenyl]allyl]tetrahydropyran-3,4,5-triol

To the residue from Step I (90 mg) in 2 mL of MeOH are added 2 drops ofMeONa (25% wt/v in MeOH). After 2h at RT, the filtrate is neutralizedwith resin Amberlite 120 (H), filtered and concentrated to dryness. Theresidue is purified on Biotage™ SNAP 10 g silica gel cartridge usingMeOH/CH₂Cl₂ (0 to 20%) in 20 column volume to afford title compound (44mg, 59%). LC-MS: m/z=827.6 (M+H⁺).

Step III: Compound 40

To(2R,3S,4R,5S,6R)-2-(hydroxymethyl)-6-[(E)-3-[3-[2-[(2R,3R,4R,5R,6R)-3,4,5-tribenzyloxy-6-(benzyloxymethyl)tetrahydropyran-2-yl]ethynyl]phenyl]allyl]tetrahydropyran-3,4,5-triol(44 mg, 0.0532 mmol) in CH₃CN (1.3 mL) is added TMSI (68 μL, 0.479mmol). The reaction mixture is stirred at RT overnight, quenched withH₂O (2 drops) and concentrated to dryness. The residue is purified byreverse phase HPLC to provide the title compound (4.2 mg, 15.7%). ¹H NMR(400 MHz, CD₃OD) δ 7.47 (s, 1H), 7.44-7.32 (m, 1H), 7.27 (dd, 2H), 6.48(d, 1H), 6.42-6.24 (m, 1H), 4.84 (d, 1H), 4.02-3.68 (m, 10H), 3.67-3.58(m, 2H), 3.57-3.48 (m, 1H), 2.63 (dd, 1H), 2.49 (dd, 1H). LC-MS:m/z=467.4 (M+H⁺).

Example 23. Preparation of Compound 41((2R,3S,4R,5S,6R)-2-(Hydroxymethyl)-6-[[4-[4-[[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]methyl]phenyl]phenyl]methyl]tetrahydropyran-3,4,5-triol)

Step I:[(2R,3S,6S)-3-Acetoxy-6-[(4-iodophenyl)methyl]-3,6-dihydro-2H-pyran-2-yl]methylAcetate (β-Isomer) and[(2R,3S,6R)-3-acetoxy-6-[(4-iodophenyl)methyl]-3,6-dihydro-2H-pyran-2-yl]methylAcetate (α-Isomer)

To a suspension of Rieke zinc (4.80 mL of 10% w/v, 7.35 mmol) in THF at0° C. is added dropwise a solution of 1-(bromomethyl)-4-iodo-benzene(1.091 g, 3.674 mmol) in THF (2 mL) over 20 min. The reaction mixture isstirred at 0° C. for 2 h and TLC revealed that most of the SM (benzylbromide) is consumed. The mixture is warmed to RT, filtered undernitrogen via a frit in a separate RBF. THF is removed under a nitrogenflow while warming the solution in a warm water bath. The dark residueis dissolved/suspended in CH₂Cl₂ (8 mL), cooled to −30° C. and[(2R,3S,4R)-3,4-diacetoxy-3,4-dihydro-2H-pyran-2-yl]methyl acetate (500mg, 1.837 mmol) is added as a solid followed by BF₃.OEt₂ (1.433 g, 1.25mL, 10.10 mmol). The final reaction mixture is warmed to 0° C. and isstirred for 45 min. TLC showed that all the starting glucal is consumed.The mixture is diluted with CH₂Cl₂ and brine. The organic fraction isisolated in Phase Separator column, dried (Na₂SO₄), filtered andconcentrated. The resulting crude mixture is purified on a Biotage™ SNAPsilica gel cartridge (50 g) using EtOAc in Hex (10 to 20%) as theeluent. A second purification is performed on the mixed fraction toafford β-isomer (187 mg) and α-isomer (282 mg). LC-MS: m/z=452.81(M+Na⁺) for β-isomer. LC-MS: m/z=452.77 (M+Na⁺) for α-isomer.

Step II:[(2R,3R,4R,5R,6R)-3,4,5-Triacetoxy-6-[(4-iodophenyl)methyl]tetrahydropyran-2-yl]methylAcetate

To a solution of[(2R,3S,6R)-3-acetoxy-6-[(4-iodophenyl)methyl]-3,6-dihydro-2H-pyran-2-yl]methylacetate (α-isomer, 282 mg, 0.616 mmol) in tert-butanol (6 mL) and water(2 mL) is added in order NMO (160 mg, 1.37 mmol), methanesulfonamide(90.0 mg, 0.946 mmol), 2,6-dimethylpyridine (75.0 μL, 0.648 mmol) andfinally OsO₄ (300 μL of 2.5% w/v, 0.0295 mmol). The reaction mixture isstirred at RT for 24 h and is monitored by LCMS. The reaction mixture ispoured in 10% NaHSO₃ and diluted with EtOAc. The organic layer isseparated, washed with water, brine, dried over Na₂CO₃, filtered andconcentrated. The resulting crude mixture is purified on Biotage™ SNAPsilica gel column (25 g) using EtOAC using Hex (10 to 60%) as theeluent. Three distinct set of fractions are isolated from which latertwo fractions are combined and per acetylated in pyridine/Ac₂O overnightat RT. Mixture is concentrated, coevaporated with toluene twice and theresulting crude mixture is purified on a Biotage™ SNAP silica gel column(10 g) using EtOAC in Hex (10 to 50%) as the eluent o afford the titlecompound (204 mg, 63.1%) which contains 5% of the 2,3 alphadiastereoisomer.

Step III:[(2R,3R,4R,5R,6R)-3,4,5-Triacetoxy-6-[[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]methyl]tetrahydropyran-2-yl]methylAcetate

To a solution of compound from Step II (112 mg, 0.204 mmol) in DMF (3mL) is added4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolane(73 mg, 0.288 mmol), potassium acetate (81 mg, 0.825 mmol) andPd(dppf)Cl₂. CH₂Cl₂ (16 mg, 0.0196 mmol). The resulting suspension isdegassed three times (house vacuum, then N₂) and stirred at 70° C. for 6h. The resulting mixture is poured in saturated aqueous NH₄Cl andextracted with Et₂O. The organic phase is washed with water (2×), brine,dried over MgSO₄, filtered and concentrated. The resulting crude mixtureis purified on a Biotage™ SNAP silica gel column (10 g) using EtOAc inHex (10 to 40%) as the eluent to afford the title compound (83 mg,72.6%).

Step IV:[(2R,3R,4R,5R,6R)-3,4,5-Triacetoxy-6-[[4-[4-[[(2R,3R,4R,5R,6R)-3,4,5-triacetoxy-6-(acetoxymethyl)tetrahydropyran-2-yl]methyl]phenyl]phenyl]methyl]tetrahydropyran-2-yl]methylAcetate

To a solution of[(2R,3R,4R,5R,6R)-3,4,5-triacetoxy-6-[(4-iodophenyl)methyl]tetrahydropyran-2-yl]methylacetate (91 mg, 0.166 mmol) and[(2R,3R,4R,5R,6R)-3,4,5-triacetoxy-6-[[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]methyl]tetrahydropyran-2-yl]methylacetate (83 mg, 0.148 mmol) in dioxane (5 mL) is added K₃PO₄ (98 mg,0.462 mmol) and finally PdCl₂(dppf).CH₂Cl₂ (10 mg). The reaction mixtureis degassed three times (house vacuum then nitrogen), stirred at 80° C.for 5 h. Final dark brown mixture is cooled to RT, diluted with 15 mLEtOAc and filtered on a pad of silica gel. The latter is washed withEtOAc (2×15 mL). The combined fractions are concentrated and the residueis purified on a Biotage™ SNAP silica gel column (10 g) using EtOAc inHex (10 to 80%) as the eluent to afford title compound (26 mg).

Step V: Compound 41

To a solution of[(2R,3R,4R,5R,6R)-3,4,5-triacetoxy-6-[[4-[4-[[(2R,3R,4R,5R,6R)-3,4,5-triacetoxy-6-(acetoxymethyl)tetrahydropyran-2-yl]methyl]phenyl]phenyl]methyl]tetrahydropyran-2-yl]methylacetate (26 mg, 0.031 mmol) in MeOH (1 mL) is added MeONa (60 μL of 0.5M, 0.030 mmol) in MeOH. The reaction mixture is stirred at RT for 12 h.Reaction mixture is quenched with acetic acid (2.0 μL, 0.035 mmol) thenconcentrated, and purified by reverse phase HPLC to afford the titlecompound (12 mg, 76.2%). ¹H NMR (400 MHz, CD₃OD) δ 7.54 (d, J=8.2 Hz,4H), 7.34 (d, J=8.2 Hz, 4H), 4.16-4.07 (m, 2H), 3.87-3.76 (m, 6H),3.76-3.61 (m, 6H), 3.07 (dd, J=14.0, 8.3 Hz, 2H), 2.94 (dd, J=14.0, 6.8Hz, 2H). LC-MS: m/z=507.46 (M+H⁺).

Example 24. Preparation of Compound 42((2R,3S,4R,5S,6R)-2-(Hydroxymethyl)-6-[1-[(2S,3S,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]triazol-4-yl]tetrahydropyran-3,4,5-triol)

Step I:[(2R,3R,4S,5S,6S)-3,4,5-Triacetoxy-6-[4-[(2R,3R,4R,5R,6R)-3,4,5-tribenzyloxy-6-(benzyloxymethyl)tetrahydropyran-2-yl]triazol-1-yl]tetrahydropyran-2-yl]methylAcetate

To a stirred mixture of the Intermediate G (76 mg, 0.1386 mmol) and[(2R,3R,4S,5S,6S)-4,5-diacetoxy-2-(acetoxymethyl)-6-azido-tetrahydropyran-3-yl]acetate(70 mg, 0.1875 mmol) in ethanol (1 mL) and water (250 μL) is added CuSO₄(11 mg, 0.0689 mmol) and sodium ascorbate (22 mg, 0.1249 mmol) in oneportion. Resultant reaction mixture is stirred for 30 min, additionalamount of ethanol (1 mL) is added, stirred at RT for 24 h, diluted withwater, extracted with methylene chloride. The combined extracts aredried, concentrated, purified on Biotage™ SNAP 12 g silica gel cartridgeusing EtOAc in Hex (15% to 50%) as eluent to afford the title compound(80 mg, 0.08677 mmol, 59.84%) as white solid. Rf=0.36 (50% EA in Hex).¹H NMR (400 MHz, CDCl₃) δ 7.64 (s, 1H), 7.40 (dd, J=7.7, 1.5 Hz, 2H),7.37-7.21 (m, 16H), 7.17-7.09 (m, 2H), 5.98 (dd, J=3.6, 2.7 Hz, 1H),5.95-5.88 (m, 2H), 5.36 (t, J=9.1 Hz, 1H), 5.32 (d, J=2.8 Hz, 1H), 4.82(d, J=11.0 Hz, 1H), 4.77 (s, 2H), 4.69-4.53 (m, 5H), 4.48 (d, J=11.0 Hz,1H), 4.35 (dd, J=12.5, 5.3 Hz, 1H), 4.03 (dd, J=12.5, 2.5 Hz, 1H), 3.96(t, J=8.7 Hz, 1H), 3.86-3.77 (m, 2H), 3.76-3.71 (m, 2H), 3.69-3.62 (m,1H), 2.19 (s, 3H), 2.07 (s, 3H), 2.05 (s, 3H), 2.05 (s, 3H).

Step II:(2R,3S,4S,5S,6S)-2-(Hydroxymethyl)-6-[4-[(2R,3R,4R,5R,6R)-3,4,5-tribenzyloxy-6-(benzyloxymethyl)tetrahydropyran-2-yl]triazol-1-yl]tetrahydropyran-3,4,5-triol

To a stirred solution of[(2R,3R,4S,5S,6S)-3,4,5-triacetoxy-6-[4-[(2R,3R,4R,5R,6R)-3,4,5-tribenzyloxy-6-(benzyloxymethyl)tetrahydropyran-2-yl]triazol-1-yl]tetrahydropyran-2-yl]methylacetate (80 mg, 0.0868 mmol) in MeOH (4 mL) is added MeONa in MeOH (185μL of 0.5 M, 0.0925 mmol). The reaction mixture is stirred at RTovernight, neutralized with acetic acid (10 μL, 0.176 mmol), ¼ of thereaction mixture (1 mL) is concentrated, freeze dried with CH₃CN-waterto afford the title compound (16 mg, 0.0191 mmol, 88.1%) as white solid.¹H NMR (400 MHz, CD₃OD) δ 8.05 (s, 1H), 7.37-7.11 (m, 20H), 5.97 (d,J=2.7 Hz, 1H), 5.24 (d, J=4.1 Hz, 1H), 4.73-4.45 (m, 10H), 4.42 (dd,J=4.0, 2.5 Hz, 1H), 4.04 (dd, J=8.5, 3.5 Hz, 1H), 3.98-3.89 (m, 2H),3.81-3.63 (m, 6H). LC-MS: m/z=754.58 (M+H⁺).

Step III: Compound 42

A solution of(2R,3S,4S,5S,6S)-2-(hydroxymethyl)-6-[4-[(2R,3R,4R,5R,6R)-3,4,5-tribenzyloxy-6-(benzyloxymethyl)tetrahydropyran-2-yl]triazol-1-yl]tetrahydropyran-3,4,5-triolin MeOH (3 mL) and acetic acid (40 μL, 0.703 mmol) is addeddihydroxypalladium (50 mg, 0.0712 mmol), stirred under hydrogen at 1atm. for 48 h, filtered through celite. The filtrate is concentrated andpurified on preparative HPLC using Phenomenex C18 Gemini AXIA 5μ, 110A21.2×75 mm 0% ACN/H₂O+0.01% TFA-To 30% ACN+0.01% TFA in 40 min-To 100%ACN in 1 min to afford the title compound (7.4 mg, 0.0169 mmol, 26%) asa half-white solid. ¹H NMR (400 MHz, CD₃OD) δ 8.18 (s, 1H), 6.03 (d,J=2.7 Hz, 1H), 5.14 (d, J=2.3 Hz, 1H), 4.72-4.67 (m, 1H), 4.48 (t, J=2.8Hz, 1H), 4.06 (dd, J=8.5, 3.5 Hz, 1H), 3.86-3.65 (m, 7H), 3.45-3.33 (m,2H).

Example 25. Preparation of Compound 43(2-[(2R,3S,4R,5S,6R)-3,4,5-Trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]-N-[4-[[2-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]acetyl]amino]phenyl]acetamide)

Step I:[(2R,3R,4R,5R,6R)-3,4,5-Triacetoxy-6-[2-oxo-2-[4-[[2-[(2R,3R,4R,5R,6R)-3,4,5-triacetoxy-6-(acetoxymethyl)tetrahydropyran-2-yl]acetyl]amino]anilino]ethyl]tetrahydropyran-2-yl]methylAcetate

To a solution of Intermediate J (128 mg, 0.328 mmol) in 3.5 mL of DMF issequentially added benzene-1,4-diamine (14.2 mg, 0.131 mmol), DIPEA (69μL, 0.394 mmol) and HATU (125 mg, 0.328 mmol) under nitrogen atmosphere.The reaction mixture is stirred at RT for 20 h, and diluted with water(10 mL). The reaction mixture is extracted by EtOAc (5×10 mL), and thecombined organic layer are washed with water (3×5 mL), 5 mL of brine,dried over Na₂SO₄, filtered, and concentrated to dryness. The residue ispurified by flash column chromatography on silica gel (0 to 20% MeOH inCH₂Cl₂) to give the title compound (116.5 mg) that is used as is for thenext step.

Step II: Compound 43

To a solution of[(2R,3R,4R,5R,6R)-3,4,5-triacetoxy-6-[2-oxo-2-[4-[[2-[(2R,3R,4R,5R,6R)-3,4,5-triacetoxy-6-(acetoxymethyl)tetrahydropyran-2-yl]acetyl]amino]anilino]ethyl]tetrahydropyran-2-yl]methylacetate (116.5 mg) in MeOH (2.3 mL) is added MeONa (68 μL of 0.5 M,0.034 mmol) in MeOH. The mixture is stirred at RT for 3 h. AcOH (24,0.034 mmol) is added and the mixture is evaporated to dryness. Theresidue is purified by preparative reverse phase HPLC to give the titlecompound (34.3 mg, 43% for last two steps). ¹H NMR (400 MHz, DMSO-D₆) δ9.76 (d, J=8.8 Hz, 2H), 7.43 (s, 4H), 4.77 (s, 2H), 4.64 (s, 2H), 4.54(s, 2H), 4.29 (s, 2H), 4.09 (m, 2H), 3.50 (s, 8H), 3.43 (s, 2H), 3.36(m, 2H), 2.51 (m, 4H).

Example 26. Preparation of Compound 44((2R,3S,4R,5S,6R)-2-(Hydroxymethyl)-6-[3-[3-[3-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]phenoxy]propoxy]phenyl]tetrahydropyran-3,4,5-triol)

To a solution of[(2R,3S,4R,5S,6R)-3-acetoxy-4,5-dihydroxy-6-(3-hydroxyphenyl)tetrahydropyran-2-yl]methylacetate (50 mg, 0.147 mmol) in DMF (1 mL) is added Cs₂CO₃ (71.81 mg,0.22 mmol) followed by 1,3-diiodopropane (10 μL, 0.088 mmol). Thereaction mixture is heated at 55° C. overnight, filtered andconcentrated. To the previous residue in MeOH (1 mL) is added catalyticMeONa (8 μL of 25% w/v in MeOH, 0.037 mmol). The reaction mixture isstirred at RT for 1h, neutralized with Amberlite IR120(H). Afterfiltration, the solvent is removed under reduced pressure and theresidue is purified by reverse phase HPLC to provide the title compound(4.8 mg, 10.8%). ¹H NMR (400 MHz, CD₃OD) δ 7.17 (t, 2H), 7.01 (s, 2H),6.92 (d, 2H), 6.76 (d, 2H), 4.84 (m, 2H), 4.32 (t, 2H), 4.09 (t, 4H),3.82-3.66 (m, 4H), 3.61 (t, 2H), 3.48 (dd, 2H), 3.44-3.31 (m, 2H),2.22-2.08 (m, 2H). LC-MS: m/z=553.4 (M+H⁺).

Example 27. Preparation of Compound 45(N-[(2R,3R,4R,5S,6R)-4,5-Dihydroxy-6-(hydroxymethyl)-2-[4-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]triazol-1-yl]tetrahydropyran-3-yl]acetamide)

To a solution of Intermediate M (15 mg, 0.0797 mmol) andN-((2R,3R,4R,5S,6R)-2-azido-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)acetamide(20 mg, 0.0812 mmol) in ethanol (0.4 mL) and water (0.1 mL) issequentially added CuSO₄ (6.0 mg, 0.0376 mmol) and sodium ascorbate (15mg, 0.0757 mmol) at RT. The reaction mixture is stirred at RT for 24 h,diluted with water and MeOH, filtered off. The filtrate is concentratedand purified on prep. HPLC on Phenomenex C18 Gemini AXIA Pack 5μ 110A21.2×250 mm 0% To 40% ACN+0.1% Formic acid in 40 min-To 100% ACN in 5min-Hold 5 min to afford the title compound (5 mg, 14%) as white solid.¹H NMR (400 MHz, CD₃OD) δ 8.10 (s, 1H), 5.66 (d, J=9.8 Hz, 1H), 5.00 (d,J=2.1 Hz, 1H), 4.38-4.35 (m, 1H), 4.13 (t, J=10.0 Hz, 1H), 3.71-3.42 (m,10H), 1.69 (s, 3H). LC-MS: m/z=435.32 (M+H⁺).

Example 28. Preparation of Compound 46 and Compound 47 (Method B)((2R,3S,4S,5S,6S)-2-(Hydroxymethyl)-6-[5-[(2S,3S,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]-2-thienyl]tetrahydropyran-3,4,5-triol)and((2R,3S,4R,5S,6R)-2-(Hydroxymethyl)-6-[5-[(2S,3S,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]-3-thienyl]tetrahydropyran-3,4,5-triol)

Step I:[(2R,3R,4S,5S,6S)-3,4,5-Tris(2,2-dimethylpropanoyloxy)-6-[5-[(2S,3S,4S,5R,6R)-3,4,5-tris(2,2-dimethylpropanoyloxy)-6-(2,2-dimethylpropanoyloxymethyl)tetrahydropyran-2-yl]-2-thienyl]tetrahydropyran-2-yl]methyl2,2-dimethylpropanoate, and[(2R,3R,4R,5R,6R)-3,4,5-tris(2,2-dimethylpropanoyloxy)-6-[5-[(2S,3S,4S,5R,6R)-3,4,5-tris(2,2-dimethylpropanoyloxy)-6-(2,2-dimethylpropanoyloxymethyl)tetrahydropyran-2-yl]-3-thienyl]tetrahydropyran-2-yl]methyl2,2-dimethylpropanoate

A solution of n-Bu₃MgLi (554 μL of 0.65 M, 0.3600 mmol) inhexane-heptane-dibutylether (8:20:3) is added to 2,5-diiodothiophene(150 mg, 0.4465 mmol) in toluene (0.5 mL) at 0° C., stirred at the sametemperature for 3.5 h (a thick precipitate is formed), a solution ofZnBr₂—LiBr in dibutyl ether (543 μL of 1.05 M, 0.57 mmol) is addeddropwise, cooling bath removed, stirred at RT for 1 h. A solution of[(2R,3R,4S,5S,6R)-6-bromo-3,4,5-tris(2,2-dimethylpropanoyloxy)tetrahydropyran-2-yl]methyl2,2-dimethylpropanoate (500 mg, 0.8628 mmol) in toluene (0.9 mL) isadded, it is placed on pre-heated oil bath at 90° C. for 4 h. It iscooled to RT and it is poured into aqueous 1 N HCl solution (10 mL),then extracted with EtOAc (3×10 mL). The combined extracts are washedwith brine, dried (Na₂SO₄), concentrated, purified on Biotage™ SNAP 50 gsilica gel cartridge using EtOAc in Hex (5% to 10%, 8CV, 10%, 5CV) aseluent to afford[(2R,3R,4S,5S,6S)-3,4,5-tris(2,2-dimethylpropanoyloxy)-6-[5-[(2S,3S,4S,5R,6R)-3,4,5-tris(2,2-dimethylpropanoyloxy)-6-(2,2-dimethylpropanoyloxymethyl)tetrahydropyran-2-yl]-2-thienyl]tetrahydropyran-2-yl]methyl2,2-dimethylpropanoate (160 mg, 0.1480 mmol, 33.1%) as white solid.Rf=0.38 (15% EA in Hex). ¹H NMR (400 MHz, CDCl₃) δ 7.23 (s, 2H), 5.87(t, J=2.6 Hz, 2H), 5.55 (t, J=9.6 Hz, 2H), 5.25 (dd, J=9.8, 3.0 Hz, 2H),5.19 (d, J=2.2 Hz, 2H), 4.30 (dd, J=12.4, 4.7 Hz, 2H), 4.09 (dd, J=12.4,1.9 Hz, 2H), 3.93 (ddd, J=9.3, 4.6, 1.8 Hz, 2H), 1.28 (s, 18H), 1.26 (s,18H), 1.16 (s, 18H), 1.15 (s, 18H) and[(2R,3R,4R,5R,6R)-3,4,5-tris(2,2-dimethylpropanoyloxy)-6-[5-[(2S,3S,4S,5R,6R)-3,4,5-tris(2,2-dimethylpropanoyloxy)-6-(2,2-dimethylpropanoyloxymethyl)tetrahydropyran-2-yl]-3-thienyl]tetrahydropyran-2-yl]methyl2,2-dimethylpropanoate (30 mg, 6.2%). Rf=0.37 (15% EA-Hex). ¹H NMR (400MHz, CDCl₃) δ 7.48 (s, 1H), 7.26 (s, 1H), 5.90 (t, J=2.6 Hz, 1H), 5.80(t, J=2.6 Hz, 1H), 5.65 (t, J=9.8 Hz, 1H), 5.56 (t, J=9.8 Hz, 1H), 5.23(s, 1H), 5.21-5.14 (m, 2H), 5.04 (s, 1H), 4.36-4.26 (m, 2H), 4.24-4.07(m, 3H), 3.88 (d, J=9.5 Hz, 1H), 1.29 (s, 9H), 1.29 (s, 9H), 1.27 (s,9H), 1.27 (s, 9H), 1.15 (s, 9H), 1.14 (s, 9H), 1.13 (s, 9H), 1.12 (s,9H).

Step II: Compound 46

To a stirred suspension of[(2R,3R,4S,5S,6S)-3,4,5-tris(2,2-dimethylpropanoyloxy)-6-[5-[(2S,3S,4S,5R,6R)-3,4,5-tris(2,2-dimethylpropanoyloxy)-6-(2,2-dimethylpropanoyloxymethyl)tetrahydropyran-2-yl]-2-thienyl]tetrahydropyran-2-yl]methyl2,2-dimethylpropanoate (92 mg, 0.0851 mmol) in MeOH (2 mL) is addedMeONa (340 μL of 0.5 M, 0.1702 mmol). It is stirred at RT 48 h, to theresultant suspension is added DOWEX 50WX4-400 until pH 4-5, suspensionbecame clear solution, filtered, concentrated, freeze dried to affordthe title compound (26.5 mg, 0.0616 mmol, 72.5%) as white fluffy solid.¹H NMR (400 MHz, CD₃OD) δ 6.94 (s, 2H), 5.12 (d, J=2.2 Hz, 2H), 4.31 (t,J=2.5 Hz, 2H), 3.83-3.64 (m, 8H), 3.56-3.47 (m, 2H).

Step III: Compound 47

To a stirred suspension of[(2R,3R,4R,5R,6R)-3,4,5-tris(2,2-dimethylpropanoyloxy)-6-[5-[(2S,3S,4S,5R,6R)-3,4,5-tris(2,2-dimethylpropanoyloxy)-6-(2,2-dimethylpropanoyloxymethyl)tetrahydropyran-2-yl]-3-thienyl]tetrahydropyran-2-yl]methyl2,2-dimethylpropanoate (33 mg, 0.0305 mmol) in MeOH (600 μL) is addedMeONa (122 μL of 0.5 M, 0.06104 mmol), stirred at RT 48 h. To theresultant solution is added DOWEX 50WX4-400 until pH 4-5, filtered,concentrated, freeze dried to afford the title compound (9.5 mg, 0.0221mmol, 72.4%) as white fluffy solid. ¹H NMR (400 MHz, CD₃OD) δ 7.29 (s,1H), 7.11 (s, 1H), 5.13 (s, 1H), 4.93 (s, 1H), 4.36 (t, J=2.8 Hz, 1H),4.32 (t, J=2.6 Hz, 1H), 3.86-3.34 (m, 10H).

Example 29. Preparation of Compound 48(2-[(2R,3S,4R,5S,6R)-3,4,5-Trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]-N-[3-[[2-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]acetyl]amino]phenyl]acetamide)

Step I:[(2R,3R,4R,5R,6R)-3,4,5-Triacetoxy-6-[2-oxo-2-[3-[[2-[(2R,3R,4R,5R,6R)-3,4,5-triacetoxy-6-(acetoxymethyl)tetrahydropyran-2-yl]acetyl]amino]anilino]ethyl]tetrahydropyran-2-yl]methylAcetate

To a solution of Intermediate J (70 mg, 0.179 mmol) in DMF (1.4 mL) areadded benzene-1,3-diamine (9.7 mg, 0.0896 mmol), 2,6-lutidine (62 μL,0.538 mmol) followed by HATU (85.2 mg, 0.224 mmol) at 0° C. The RM isstirred at RT for 3h, diluted with EtOAc, washed with H₂O, brine. Theorganic phase is dried over Na₂SO₄, filtered and concentrated. Theresidue is purified on Biotage™ SNAP 10 g silica gel cartridge usingEtOAc in Hex (0 to 15%) in 25 column volume to afford the title compound(58 mg, 75.8%) as colorless oil. LC-MS: m/z=853.5 (M+H⁺).

Step II: Compound 48

To a solution of[(2R,3R,4R,5R,6R)-3,4,5-triacetoxy-6-[2-oxo-2-[3-[[2-[(2R,3R,4R,5R,6R)-3,4,5-triacetoxy-6-(acetoxymethyl)tetrahydropyran-2-yl]acetyl]amino]anilino]ethyl]tetrahydropyran-2-yl]methylacetate (58 mg, 0.068 mmol) in MeOH (1.4 mL) is added 2 drops 25% w/vMeONa in MeOH. After stirring for 1h, the reaction mixture isneutralized with Amberlite IR120(H). After filtration, washing withMeOH, the solvent is removed under reduced pressure and the residuepurified by reverse phase HPLC to provide the title compound (11 mg,23.3%). ¹H NMR (400 MHz, CD₃OD) δ 7.84 (d, 1H), 7.37-7.26 (m, 2H),7.26-7.11 (m, 1H), 4.39-4.29 (m, 2H), 3.87 (dd, 2H), 3.79-3.64 (m, 8H),3.64-3.55 (m, 2H), 2.71 (qd, 4H). LC-MS: m/z=517.4 (M+H⁺).

Example 30. Preparation of Compound 49(2-[(2R,3S,4R,5S,6R)-3,4,5-Trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]-N-[[4-[[[2-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]acetyl]amino]methyl]phenyl]methyl]acetamide)

Compound 49 is prepared according to a similar procedure as described inExample 29 but using Intermediate J and[4-(aminomethyl)phenyl]methanamine instead of benzene-1,3-diamine asreagents. ¹H NMR (400 MHz, CD₃OD) δ 7.24 (s, 4H), 4.46-4.15 (m, 6H),3.81 (dt, 2H), 3.68 (ddd, 8H), 3.60-3.49 (m, 2H), 2.75-2.40 (m, 4H).LC-MS: m/z=545.4 (M+H⁺).

Example 31. Preparation of Compound 50((2R,3S,4R,5S,6R)-2-(Hydroxymethyl)-6-[3-[4-[3-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]phenyl]buta-1,3-diynyl]phenyl]tetrahydropyran-3,4,5-triol)

A solution of methyl 4-iodobenzoate (37 mg, 0.141 mmol), CuI (4.8 mg,0.025 mmol) and Pd(dppf)Cl₂—CH₂Cl₂ (19 mg, 0.0233 mmol) in DMF (1.0 mL)is degassed (vacuum/N₂ flush), to this is added triethylamine (70.1 mg,97 μL, 0.6924 mmol) and Intermediate K (35 mg, 0.115 mmol) undernitrogen atmosphere, reaction mixture is stirred at RT overnight,directly loaded onto C-18 samplet, purified on Biotage™ SNAP 25 g C18silica gel cartridge using CH₃CN-water (5% to 50%) as eluent to affordthe title compound (3.0 mg). ¹H NMR (400 MHz, CD₃OD) δ 7.62 (s, 2H),7.51 (d, J=7.4 Hz, 2H), 7.41 (d, J=7.7 Hz, 2H), 7.35 (t, J=7.7 Hz, 2H),4.88 (d, J=4.5 Hz, 2H), 4.29 (dd, J=4.4, 3.2 Hz, 2H), 3.84 (dd, J=11.9,6.9 Hz, 2H), 3.76 (dd, J=11.9, 3.1 Hz, 2H), 3.71 (t, J=7.2 Hz, 2H), 3.56(dd, J=7.4, 3.1 Hz, 2H), 3.51-3.43 (m, 2H). LC-MS: m/z=527.51 (M+H⁺).

Example 32. Preparation of Compound 51(1,3-Bis(3-((2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)phenyl)urea)

Step I:3-((2R,3R,4R,5R,6R)-3,4,5-Triacetoxy-6-(acetoxymethyl)tetrahydro-2H-pyran-2-yl)benzoicAcid

To a suspension of Intermediate D (165 mg, 0.551 mmol) in 1.6 mL ofCH₂Cl₂ is sequentially added pyridine (312 μL, 3.86 mmol), DMAP (6.7 mg,0.055 mmol) and Ac₂O (312 μL, 3.31 mmol) under nitrogen atmosphere. Thereaction mixture is stirred at RT for 20 h and diluted with 2M HCl (0.5mL). The organic layer is dried over Na₂SO₄, filtered, and concentratedto dryness. The residue is purified by flash column chromatography onsilica gel (0 to 20% MeOH in CH₂Cl₂) to give the raw product. Theproduct is dissolved in 5 mL of CH₂Cl₂ and washed with 1M HCl (1 mL).The organic layer is dried over Na₂SO₄, filtered, and concentrated todryness to give the title compound (242.2 mg, 97%).

Step II:[(2R,3R,4R,5R,6R)-3,4,5-Triacetoxy-6-[3-[[3-[(2R,3R,4R,5R,6R)-3,4,5-triacetoxy-6-(acetoxymethyl)tetrahydropyran-2-yl]phenyl]carbamoylamino]phenyl]tetrahydropyran-2-yl]methylAcetate

To a solution of3-((2R,3R,4R,5R,6R)-3,4,5-triacetoxy-6-(acetoxymethyl)tetrahydro-2H-pyran-2-yl)benzoicacid (242 mg, 0.535 mmol) in 7.3 mL of tert-butanol is sequentiallyadded TEA (112 μL, 0.802 mmol) and diphenylphosphoryl azide (162 mg,0.588 mmol) under nitrogen atmosphere. The reaction mixture is heated toreflux for 4 h, cooled to RT, and concentrated to dryness. The residueis purified by flash column chromatography on silica gel (2 to 20% MeOHin CH₂Cl₂) to give the title compound (113 mg, 48%).

Step III: Compound 51

To a solution of[(2R,3R,4R,5R,6R)-3,4,5-triacetoxy-6-[3-[[3-[(2R,3R,4R,5R,6R)-3,4,5-triacetoxy-6-(acetoxymethyl)tetrahydropyran-2-yl]phenyl]carbamoylamino]phenyl]tetrahydropyran-2-yl]methylacetate in MeOH (2 mL) is added MeONa (64 μL of 0.5 M, 0.032 mmol) inMeOH. The mixture is stirred at RT overnight and filtered on a 1 g SCX-2SPE cartridge. The filtrate is evaporated to dryness and the residue ispurified by preparative HPLC to give the title compound (38 mg, 49%). ¹HNMR (400 MHz, DMSO-D₆) δ 8.66 (s, 2H), 7.44 (s, 2H), 7.37 (d, J=8.1 Hz,2H), 7.21 (t, J=7.9 Hz, 2H), 7.00 (d, J=7.7 Hz, 2H), 4.75 (m, 4H), 4.64(m, 4H), 4.54 (t, J=5.8 Hz, 2H), 4.03 (m, 2H), 3.60 (m, 4H), 3.54 (m,2H), 3.45-3.36 (m, 4H).

Example 33. Preparation of Compound 52((2R,3S,4R,5S,6R)-2-(Hydroxymethyl)-6-[3-[4-[3-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]phenyl]butyl]phenyl]tetrahydropyran-3,4,5-triol)

A mixture of(2R,3S,4R,5S,6R)-2-(hydroxymethyl)-6-[3-[4-[3-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]phenyl]buta-1,3-diynyl]phenyl]tetrahydropyran-3,4,5-triol(Compound 50, 5 mg, 0.009 mmol) and 10% Pd on C, wet, Degussa (10 mg,0.0094 mmol) in MeOH (3 mL) is hydrogenated at 40 psi for 4 h, filteredoff catalyst, concentrated. This material is dissolved inwater-acetonitrile and freeze dried to afford title compound (4.5 mg,86%) as white solid. ¹H NMR (400 MHz, CD₃OD) δ 7.21 (s, 2H), 7.17 (d,J=4.9 Hz, 4H), 6.99 (t, J=3.7 Hz, 2H), 4.86 (d, J=3.3 Hz, 2H), 4.35 (t,J=3.3 Hz, 2H), 3.71 (d, J=4.6 Hz, 4H), 3.63 (t, J=8.2 Hz, 2H), 3.47 (dd,J=8.2, 3.1 Hz, 2H), 3.41-3.32 (m, 2H), 2.62-2.47 (m, 4H), 1.63-1.48 (m,4H). LC-MS: m/z=535.53 (M+H⁺).

Example 34. Preparation of Compound 53 (Method C)(2R,3S,4R,5S,6R)-2-(Hydroxymethyl)-6-[2-[4-[2-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]ethynyl]phenyl]ethynyl]tetrahydropyran-3,4,5-triol

Step I:[(2R,3R,4R,5R,6R)-6-[2-[4-[2-[(2R,3R,4R,5R,6R)-6-(Hydroxymethyl)-3,4,5-tris(triisopropylsilyloxy)tetrahydropyran-2-yl]ethynyl]phenyl]ethynyl]-3,4,5-tris(triisopropylsilyloxy)tetrahydropyran-2-yl]methanol

To a degassed (vacuum-nitrogen flush, 30 min) solution of Intermediate L(836 mg, 1.15 mmol), 1,4-diiodobenzene (180 mg, 0.545 mmol),PdCl₂(dppf). CH₂Cl₂ (44.6 mg, 0.0546 mmol) and CuI (10.4 mg, 0.0546mmol) in DMF (8 mL) under nitrogen atmosphere are added DBU (653 μL,4.37 mmol) and degassed water (29.5 mg, 30 μL, 1.64 mmol). The reactionmixture is slowly heated at 50° C. for 4 h, and then increased to 55° C.and heated for 16 h. Then additional amount of water (10 uL) is added,continued to heat for 6 h, cooled to rt and left it overnight. Thereaction mixture is diluted with water (25 mL), extracted with EtOAc(3×25 mL), combined extracts are washed with brine, dried (Na₂SO₄) andconcentrated. The residue is purified on Biotage™ SNAP 100 g silica gelcartridge using EtOAc in Hex (0% to 20%, 8 CV; 20% 3 CV) as eluent toafford title compound (511 mg, 0.368 mmol, 68%) as half-white solid. ¹HNMR (400 MHz, CDCl₃) δ 7.37 (d, J=0.9 Hz, 4H), 4.90 (d, J=8.9 Hz, 2H),4.44 (d, J=9.1 Hz, 2H), 4.20 (brs, 2H), 4.14-4.03 (m, 2H), 4.01-3.83 (m,4H), 3.67-3.51 (m, 2H), 2.1-2.0 (m, 2H), 1.33-0.77 (m, 126H).

Step II, Compound 53

A solution of[(2R,3R,4R,5R,6R)-6-[2-[4-[2-[(2R,3R,4R,5R,6R)-6-(hydroxymethyl)-3,4,5-tris(triisopropylsilyloxy)tetrahydropyran-2-yl]ethynyl]phenyl]ethynyl]-3,4,5-tris(triisopropylsilyloxy)tetrahydropyran-2-yl]methanol(471 mg, 0.339 mmol) in THF (16.0 mL), TFA (8.0 mL) and water (8.0 mL)is heated to reflux for (bath temperature 75° C.) 30 h and concentrated.The residue is purified on Biotage™ SNAP 25 g C18 silica gel cartridgeusing CH₃CN in water (0% to 50%, 10 CV) as eluent to afford titlecompound (146 mg, 0.311 mmol, 88%) as half-white solid. ¹H NMR (400 MHz,CD₃OD) δ 7.43 (s, 4H), 4.85 (d, J=2.1 Hz, 2H), 3.98 (dd, J=3.1, 2.2 Hz,2H), 3.92-3.83 (m, 4H), 3.81-3.68 (m, 4H), 3.61 (t, J=9.5 Hz, 2H).

Alternate Synthesis of Compound 53 (Method C)

Step I:[(2R,3R,4R,5R,6R)-6-[2-[4-[2-[(2R,3R,4R,5R,6R)-6-(hydroxymethyl)-3,4,5-tris(triisopropylsilyloxy)tetrahydropyran-2-yl]ethynyl]phenyl]ethynyl]-3,4,5-tris(triisopropylsilyloxy)tetrahydropyran-2-yl]methanol

Intermediate P (118 mg, 0.18 mmol) and 1,4-diiodobenzene (28 mg, 0.083mmol) are charged in a vial, dissolved in DMF (3 mL), degassed, then CuI(7.8 mg, 0.041 mmol) and Pd(dppf)Cl₂— CH₂Cl₂ (8.4 mg, 0.010 mmol) areadded, degassed again, then Et₃N (57 μL, 0.41 mmol) is added and thereaction mixture is capped and heated at 40° C. for 4 h. The reactionmixture is diluted with EtOAc (10 mL), washed sequentially withsaturated NH₄Cl, H₂O, brine (5 mL each), dried over Na₂SO₄, filtered andconcentrated then purified on Biotage™ SNAP 10 g silica gel cartridgeusing a gradient of EtOAc in Hex (0-20%). Pure fractions are combinedand concentrated to afford title compound (60 mg, 0.043 mmol, 53% yield)as a colorless foam.

Step II, Compound 53

To a solution of[(2R,3R,4R,5R,6R)-6-[2-[4-[2-[(2R,3R,4R,5R,6R)-6-(hydroxymethyl)-3,4,5-tris(triisopropylsilyloxy)tetrahydropyran-2-yl]ethynyl]phenyl]ethynyl]-3,4,5-tris(triisopropylsilyloxy)-tetrahydropyran-2-yl]methanol(100 mg, 0.072 mmol) in THF (1 mL) is added a THF solution oftetrabutylammonium fluoride (575 μL of 1 M, 0.58 mmol). After stirringfor 3.5 h, the reaction mixture is concentrated and purified byreverse-phase flash chromatography on Biotage™ SNAP 12 g C18 silica gelcartridge using a gradient of MeCN in H₂O (0-50%). The combinedfractions containing desired compound are concentrated, the residue isredissolved in MeCN/H₂O (20% MeCN), and freeze-dried to provide 52 mg ofwhite solid.

Alternate Synthesis of Compound 53 (Method D)

To a solution of Intermediate M (130 mg, 0.6908 mmol) in DMF (4 mL) areadded 1,4-diiodobenzene (102.9 mg, 0.3119 mmol), PdCl₂(dppf)-CH₂Cl₂(13.1 mg, 0.0160 mmol), CuI (11.9 mg, 0.0625 mmol) and DIPEA (163 μL,0.936 mmol). The mixture is stirred at 60° C. for 5 h under nitrogen.After removal of the solvent under reduced pressure, the residue ispurified using prep-HPLC to obtain the title compound (52 mg, 0.115mmol, 37%) as a white solid. LC-MS: m/z=451.5 (M+H⁺).

Alternate Synthesis of Compound 53 (Method E)

Step I:(2R,2′R,3S,3′S,6S,6′S)-(1,4-phenylenebis(ethyne-2,1-diyl))bis(2-(acetoxymethyl)-3,6-dihydro-2H-pyran-6,3-diyl)diacetate

In a 1.0 L 3-neck RBF equipped with a magnetic stirrer and a thermometeris dissolvedtrimethyl-[2-[4-(2-trimethylsilylethynyl)phenyl]ethynyl]silane (25.00 g,92.42 mmol) and Indium Triflate (2.597 g, 4.621 mmol) in CH₂Cl₂ (125mL). The mixture is stirred at RT for 10 min then[(2R,3S,4R)-3,4-diacetoxy-3,4-dihydro-2H-pyran-2-yl]methyl acetate(100.7 g, 369.7 mmol) dissolved in 100 mL of CH₂Cl₂ is added dropwiseover 2 h to the reaction mixture via addition funnel keeping thereaction temperature below 25° C. At the end of the addition thereaction mixture is stirred at RT for an additional 18 h. The reactionis quenched with saturated aqueous NaHCO₃ (250 mL) and the layers areseparated. The aqueous layer is back extracted once with 150 mL ofCH₂Cl₂ and the combined organic layers are washed with water (250 mL),dried over MgSO₄ and filtered. Silica gel (500 mL) is added to theorganic phase and the mixture is evaporated to dryness under reducedpressure. The crude mixture is purified on a large pad of silica geleluting with Hex/EtOAc (10 to 50%). Fractions containing the desiredproduct are combined, concentrated to 300 mL then heptane (500 mL) andMeOH (500 mL) are added. The mixture is concentrated until a solid isformed and the latter is filtered to afford 31.0 g (58%) of the titlecompound.

Step II:(2R,2′R,3S,3′S,4R,4′R,5S,5′S,6R,6′R)-(1,4-phenylenebis(ethyne-2,1-diyl))bis(2-(acetoxymethyl)-4,5-dihydroxytetrahydro-2H-pyran-6,3-diyl)diacetate

In a 5 L 3 necks RBF equipped with a mechanical stirrer, N₂ inlet and atemperature probe is charged with K₂CO₃ (150.6 g, 1.090 mol),methanesulfonamide (51.83 g, 544.9 mmol), K₃[Fe(CN)₆] (358.9 g, 1.090mol), 2-methylpropan-2-ol (1.000 L) and water (1.250 L).1,4-bis[(S)-[(5S)-5-ethylquinuclidin-2-yl]-(6-methoxy-4-quinolyl)methoxy]phthalazine(2.830 g, 3.633 mmol) is then added followed by K₂O₄Os (334.6 mg, 0.9082mmol) and the mixture is stirred at RT for 15 min. To this mixture isadded[(2R,3S,6S)-3-acetoxy-6-[2-[4-[2-[(2R,3S,6S)-3-acetoxy-2-(acetoxymethyl)-3,6-dihydro-2H-pyran-6-yl]ethynyl]phenyl]ethynyl]-3,6-dihydro-2H-pyran-2-yl]methylacetate from Step I (50.00 g, 90.82 mmol) dissolved in EtOAc (250.0 mL)dropwise via an addition funnel over 10 min. The resulting reactionmixture (brown-orange+quite homogeneous, no chunks) is stirred (300 rpm)at RT for 18 h. An extra amount of K₂O₄Os is added (335 mg, 0.908 mmol)and the reaction mixture is stirred for 24 h. The last procedure isrepeated twice for a total amount of 1.340 g of K₂O₄Os and a reactiontime of 96 h. The reaction mixture is quenched with Na₂SO₃ (233.9 g in250 mL of water), stirred at RT for 60 min, filtered through a pad ofcelite. The organic layer is separated and concentrated to afford 76.74g of crude compound as brown gum.

The crude product and DMAP (1.110 g, 9.082 mmol) are dissolved inpyridine (250.0 mL). The resulting solution is cooled in ice/water bathand Ac₂O (51.4 mL, 545 mmol) is added dropwise over 10 minutes keepingtemperature below 10° C. The reaction mixture is then stirred at RT for1.5 h. After stirring at RT for 1 h, 25 mL of extra Ac₂O is added andthe mixture is stirred for an additional 4 h. The resulting reactionmixture is diluted with CH₂Cl₂ (250 mL) and water (250 mL), stirred for15 min, then 2N HCl (2.25 L) is added and stirred for 5 min withice/water bath to control exotherm. The aqueous solution is separatedand back-extracted with CH₂Cl₂ (2×250 mL) and the combined organicextracts are washed once again with 2N HCl (250 mL, added brine to helpseparation), dried over Na₂SO₄ then concentrated to afford 52.3 g ofcrude brown foamy solid. The residue is suspended in EtOH (262 mL),stirred under reflux until complete dissolution and then slowly cooleddown to RT then at 0° C. for 30 min. The resulting precipitate isfiltered, washed with cold EtOH to afford after drying in vacuum oven at35° C. for 48 hrs, 43.0 g of a beige powder (77% yield over two steps).

Step III: Compound 53

A 1 L RBF is charged[(2R,3R,4R,5R,6R)-3,4,5-triacetoxy-6-[2-[4-[2-[(2R,3R,4R,5R,6R)-3,4,5-triacetoxy-6-(acetoxymethyl)tetrahydropyran-2-yl]ethynyl]phenyl]ethynyl]tetrahydropyran-2-yl]methylacetate from Step II (11.55 g, 14.68 mmol), EtOH (347 mL) is added andthe mixture is stirred at RT for 5 minutes. MeONa (327 μL of 25% w/w,1.47 mmol) is added and the reaction mixture is stirred at RT for 4days. Reaction is incomplete and MeOH (120 mL) and additional MeONa (981μL of 25% w/w, 4.41 mmol) is added. Final mixture is stirred for 2 days,filtered and the resulting solid is washed with EtOH. The mother liquorsare neutralized through a 27 g Dowex 50W4 (prewash with EtOH) column.The solid is then swished for 15 min in the neutralized mother liquors.The slurry is then concentrated to dryness to afford 10 g of off-whitesolid. The latter is poured in EtOH (165 mL) and stirred at 70° C. for 1h. The slurry is slowly cooled down to RT then cooled and stirred at 0°C. for 30 min. Filtration and washing twice with cold EtOH (2×10 mL) andonce with Heptane (10 mL) afforded 6.70 g of an off-white solid. Dryingin a vacuum oven at 50° C. for 2 days afforded the title compound as anoff-white solid (6.37 g, 95%)

Example 35. Preparation of Compound 54(2R,3S,4R,5S,6R)-2-(hydroxymethyl)-6-[4-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]buta-1,3-diynyl]tetrahydropyran-3,4,5-triol

To a solution of Intermediate M (46 mg, 0.2444 mmol) in DMF (3 mL) areadded PdCl₂(dppf)-CH₂Cl₂ (10 mg, 0.0123 mmol), CuI (9.3 mg, 0.0488 mmol)and DIPEA (85 μL, 0.488 mmol). The mixture is stirred at 50° C.overnight under nitrogen. After removal of the solvent under reducedpressure, the residue is purified on prep-HPLC to obtain the titlecompound (6 mg, 0.0137 mmol, 6%) as a white solid. ¹H NMR (400 MHz,CD₃OD) δ 4.73 (d, 2H), 3.91-3.88 (m, 2H), 3.83 (m, 2H), 3.77-3.68 (m,4H), 3.66-3.60 (m, 2H), 3.57 (m, 2H). LC-MS: m/z=374.5 (M+H⁺).

Example 36. Preparation of Compound 55(2R,3S,4R,5S,6R)-2-(Hydroxymethyl)-6-[(E)-3-[4-[2-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]ethynyl]phenyl]allyl]tetrahydropyran-3,4,5-triol

Step I:[(2R,3R,4R,5R,6R)-3,4,5-Triacetoxy-6-[(E)-3-(4-iodophenyl)allyl]tetrahydropyran-2-yl]methylAcetate

To a solution of Intermediate H (300 mg, 0.8057 mmol) in DMF (8.0 mL)are added 1,4-diiodobenzene (265.8 mg, 0.806 mmol), palladium acetate(18.09 mg, 0.081 mmol), tetrabutylammonium bromide (259.7 mg, 0.81 mmol)and sodium bicarbonate (203.0 mg, 2.42 mmol). The reaction mixture isheated at 85° C. overnight under N₂. Water is added to reaction mixture,extracted with EtOAc (2×). Combined extracts are washed with brine/water(twice, 1:1, v/v) and dried over MgSO₄, filtered and concentrated underreduced pressure. The dark crude oil is purified on Biotage™ SNAP 25 gsilica gel using EtOAc in Hex (0 to 45% in 20 CV; 45% in 2 CV) to affordtitle compound (260 mg, 0.453 mmol, 56%) as a yellowish oil. ¹H NMR (400MHz, CD₃OD) δ 7.64 (d, J=8.4 Hz, 2H), 7.18 (d, J=8.4 Hz, 2H), 6.50 (d,J=15.9 Hz, 1H), 6.32-6.22 (m, 1H), 5.35 (dd, J=8.7, 3.3 Hz, 1H), 5.24(t, J=3.4 Hz, 1H), 5.15 (t, J=8.4 Hz, 1H), 4.40-4.31 (m, 1H), 4.15-4.00(m, 5H), 2.78-2.53 (m, 2H), 2.10 (s, 4H), 2.07 (s, 3H), 2.01 (d, J=1.6Hz, 5H), 1.90 (s, 3H), 1.24 (t, J=7.1 Hz, 2H). LC-MS: m/z=575.35 (M+H+)

Step II:[(2R,3R,4R,5R,6R)-3,4,5-Triacetoxy-6-[(E)-3-[4-[2-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]ethynyl]phenyl]allyl]tetrahydropyran-2-yl]methylAcetate

A solution of[(2R,3R,4R,5R,6R)-3,4,5-triacetoxy-6-[(E)-3-(4-iodophenyl)allyl]tetrahydropyran-2-yl]methylacetate from Step I (130 mg, 0.226 mmol), CuI (9.4 mg, 0.049 mmol) andPdCl₂(dppf)-CH₂Cl₂ (37.3 mg, 0.046 mmol) in DMF (4.2 mL) is degassed(vacuum/N₂ flush). To this are added triethylamine (137.4 mg, 189.3 μL,1.358 mmol) and Intermediate M (42.6 mg, 0.226 mmol) under nitrogenatmosphere. The reaction mixture is stirred at RT overnight. Reactionmixture is quenched with water and extracted with EtOAc (2×). Combinedorganic extracts are washed with diluted NH₄Cl, and brine/water (twice,1:1, v/v), and dried over MgSO₄, filtered and concentrated under reducedpressure to afford title compound (143 mg) and it is used as such in thenext step. LC-MS: m/z=635.52 (M+H⁺).

Step III, Compound 55

To a stirred solution of[(2R,3R,4R,5R,6R)-3,4,5-triacetoxy-6-[(E)-3-[4-[2-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]ethynyl]phenyl]allyl]tetrahydropyran-2-yl]methylacetate from step II (143 mg, 0.225 mmol) in MeOH (2.9 mL) is addedMeONa in MeOH (225 μL of 0.5 M, 0.113 mmol). The reaction mixture isstirred at RT overnight. The reaction mixture is passed through ionexchange resin Isolute SCX-2 (1 g column) and washed with dry MeOH. Thecombined filtrate is concentrated to dryness and the residue is purifiedon prep. HPLC to afford the title compound (22.3 mg, 20%) as a whitefluffy solid. ¹H NMR (400 MHz, CD₃OD) δ 7.41 (s, 4H), 6.55 (d, J=15.9Hz, 1H), 6.47-6.35 (m, 1H), 4.06-4.00 (m, 2H), 3.96 (dd, J=9.3, 3.3 Hz,1H), 3.90 (dd, J=11.5, 2.1 Hz, 1H), 3.87-3.73 (m, 6H), 3.71-3.63 (m,2H), 3.57 (ddd, J=9.0, 6.1, 3.2 Hz, 1H), 2.76-2.65 (m, 1H), 2.60-2.49(m, 1H).

Example 37. Preparation of Compound 56 (Method A)(2R,3S,4R,5S,6R)-2-(Hydroxymethyl)-6-[2-[6-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]-2-naphthyl]ethynyl]tetrahydropyran-3,4,5-triol

Step I:[(2R,3S,6S)-3-Acetoxy-6-(6-hydroxy-2-naphthyl)-3,6-dihydro-2H-pyran-2-yl]methylAcetate

Acetonitrile (10 mL) is added to a mixture of[(2R,3S,4R)-3,4-diacetoxy-3,4-dihydro-2H-pyran-2-yl]methyl acetate (1.49g, 5.47 mmol), (6-hydroxy-2-naphthyl)boronic acid (1.0 g, 5.32 mmol) anddiacetoxypalladium (119 mg, 0.53 mmol). The reaction mixture is stirredovernight at RT under N₂, then filtered through silica cartridge (5 g),and rinsed with EtOAc. Combined filtrates are concentrated, and purifiedon Biotage™ SNAP 50 g silica gel cartridge using a gradient of EtOAc inHex (0-50%). Pure fractions are combined and concentrated to providetitle compound (935 mg, 2.62 mmol, 49% yield) as an off-white foamysolid. ¹H NMR (400 MHz, CDCl₃) δ 7.80-7.72 (m, 2H), 7.70 (d, J=8.5 Hz,1H), 7.51 (dd, J=8.5, 1.7 Hz, 1H), 7.18-7.08 (m, 2H), 6.28 (ddd, J=10.4,3.1, 1.6 Hz, 1H), 6.13-6.00 (m, 1H), 5.49-5.42 (m, 1H), 5.36 (ddd,J=7.4, 4.1, 2.1 Hz, 1H), 4.28 (dd, J=12.0, 5.8 Hz, 1H), 4.09 (dd,J=12.0, 3.0 Hz, 1H), 3.86 (ddd, J=7.8, 5.8, 3.0 Hz, 1H), 2.10 (s, 3H),2.08 (s, 3H). LC-MS: m/z=357.34 (M+H⁺).

Step II:[(2R,3S,4R,5S,6R)-3-Acetoxy-4,5-dihydroxy-6-(6-hydroxy-2-naphthyl)tetrahydropyran-2-yl]methylAcetate

To a suspension of[(2R,3S,6S)-3-acetoxy-6-(6-hydroxy-2-naphthyl)-3,6-dihydro-2H-pyran-2-yl]methylacetate (930 mg, 2.61 mmol) in THF (5.6 mL)/water (3.7 mL) are addedOsO₄ (2.5% w/w in t-BuOH, 980 μL, 0.078 mmol) followed by4-methyl-4-oxido-morpholin-4-ium (917 mg, 7.83 mmol). After stirring for23 h, more OsO₄ solution is added (300 μL) and stirred another 18 h,then 1M aqueous Na₂S₂O₃ solution (6 mL), EtOAc (40 mL), and H₂O (10 mL)are added. Organic layer is separated, aqueous layer is extracted withEtOAc (2×20 mL). Combined organic extracts are washed with brine (20mL), dried over Na₂SO₄, filtered and concentrated to provide crudeproduct as a dark brown foamy solid, which is purified on Biotage™ SNAP50 g silica gel cartridge using a gradient of ^(i)PrOH in CH₂Cl₂(0-10%). Combined pure fractions are concentrated to provide titlecompound (281 mg, 0.72 mmol, 27% yield) as an off-white solid. ¹H NMR(400 MHz, CD₃OD) δ 7.77 (s, 1H), 7.73 (d, J=8.7 Hz, 1H), 7.67 (d, J=8.6Hz, 1H), 7.48 (dd, J=8.6, 1.7 Hz, 1H), 7.12-7.04 (m, 2H), 5.10 (dd,J=6.3, 5.3 Hz, 1H), 5.03 (d, J=6.2 Hz, 1H), 4.74 (dd, J=12.1, 8.1 Hz,1H), 4.31 (dd, J=6.2, 3.2 Hz, 1H), 4.12 (dd, J=12.1, 3.2 Hz, 1H),3.95-3.84 (m, 2H), 2.09 (s, 3H), 2.06 (s, 3H). LC-MS: m/z=413.34(M+Na⁺).

Step III:[(2R,3S,4R,5S,6R)-3-Acetoxy-4,5-dihydroxy-6-[6-(trifluoromethylsulfonyloxy)-2-naphthyl]tetrahydropyran-2-yl]methylAcetate

To a suspension of[(2R,3S,4R,5S,6R)-3-acetoxy-4,5-dihydroxy-6-(6-hydroxy-2-naphthyl)tetrahydropyran-2-yl]methylacetate (235 mg, 0.60 mmol) in CH₂Cl₂ (6 mL) is added Et₃N (168 μL, 1.21mmol),1,1,1-trifluoro-N-phenyl-N-(trifluoromethylsulfonyl)-methanesulfonamide(266 mg, 0.7446 mmol), and CH₂Cl₂ (4 mL). After stirring for 4 days, thereaction mixture is concentrated. Purified on Biotage™ SNAP 25 g silicagel cartridge, using a gradient of MeOH in CH₂Cl₂ (0-10%). Fractionscontaining product are combined, concentrated then purified again onBiotage™ SNAP 25 g silica gel cartridge, using a gradient of EtOAc inHex (50-80%). Pure fractions are combined and concentrated to affordtitle compound (214 mg, 0.41 mmol, 68% yield) which is obtained as ayellowish waxy solid. ¹H NMR (400 MHz, CD₃OD) δ 8.06 (d, J=9.1 Hz, 1H),8.02 (s, 1H), 8.00 (d, J=8.6 Hz, 1H), 7.91 (d, J=2.5 Hz, 1H), 7.74 (dd,J=8.6, 1.6 Hz, 1H), 7.47 (dd, J=9.0, 2.5 Hz, 1H), 5.15-5.00 (m, 2H),4.24-4.07 (m, 2H), 4.02-3.97 (m, 1H), 3.95 (dd, J=5.4, 3.1 Hz, 1H), 2.12(s, 3H), 2.03 (s, 3H). LC-MS: m/z=545.36 (M+Na⁺).

Step IV:[(2R,3R,4R,5R,6R)-3,4,5-Triacetoxy-6-[6-(trifluoromethylsulfonyloxy)-2-naphthyl]tetrahydropyran-2-yl]methylAcetate

To a solution of[(2R,3S,4R,5S,6R)-3-acetoxy-4,5-dihydroxy-6-[6-(trifluoro-methylsulfonyloxy)-2-naphthyl]tetrahydropyran-2-yl]methylacetate (214 mg, 0.41 mmol) in CH₂Cl₂ (2.2 mL) are added pyridine (100μL, 1.24 mmol), DMAP (2.5 mg, 0.0208 mmol) and Ac₂O (97 μL, 1.03 mmol).After stirring for 2 h, the reaction mixture is diluted with CH₂Cl₂ (5mL), aqueous 1N HCl (5 mL) is added, the layers are separated. Aqueouslayer is extracted with CH₂Cl₂ (2×5 mL). Combined organic extracts areconcentrated, redissolved in CH₂Cl₂, treated with prewashed Dowex50WX4-400 resin, filtered and rinsed with portions of CH₂Cl₂. Combinedfiltrates are concentrated to provide title compound (232 mg, 0.38 mmol,93% yield) as an off-white foamy solid. ¹H NMR (400 MHz, CDCl₃) δ 8.05(s, 1H), 7.99 (d, J=9.0 Hz, 1H), 7.95 (d, J=8.7 Hz, 1H), 7.77 (d, J=2.4Hz, 1H), 7.72 (dd, J=8.5, 1.5 Hz, 1H), 7.43 (dd, J=9.0, 2.5 Hz, 1H),6.10 (t, J=3.3 Hz, 1H), 5.37 (t, J=8.6 Hz, 1H), 5.27 (d, J=3.2 Hz, 1H),5.22 (dd, J=8.9, 3.1 Hz, 1H), 4.44 (dd, J=12.2, 6.3 Hz, 1H), 4.15 (dd,J=12.1, 2.8 Hz, 1H), 3.83-3.69 (m, 1H), 2.17 (s, 3H), 2.14 (s, 3H), 2.10(s, 3H), 2.02 (s, 3H). LC-MS: m/z=607.41 (M+H⁺).

Step V:[(2R,3R,4R,5R,6R)-3,4,5-Triacetoxy-6-[6-[2-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]ethynyl]-2-naphthyl]tetrahydropyran-2-yl]methylAcetate

To a mixture of[(2R,3R,4R,5R,6R)-3,4,5-Triacetoxy-6-[6-(trifluoromethylsulfonyloxy)-2-naphthyl]tetrahydropyran-2-yl]methylacetate from step IV (45 mg, 0.0742 mmol), Intermediate M (25 mg, 0.13mmol and CuI (6.3 mg, 0.033 mmol) in a microwave vial are added DMF (500μL) and Et₃N (52 μL, 0.37 mmol). The reaction mixture is degassed(house-vac then N₂, 3×), then PdCl₂(dppf)-CH₂Cl₂ (8 mg, 0.0098 mmol) isadded, degassed again, capped and stirred at 80° C. overnight. Thereaction mixture is cooled-down to RT and diluted with EtOAc (2 mL) andH₂O (1 mL), filtered through celite and rinsed with EtOAc (4×0.5 mL).Filtrate is diluted with H₂O and brine (1 mL) and EtOAc. Layers areseparated. Organic layer is washed with H₂O, brine, dried over Na₂SO₄,filtered and concentrated to provide crude product. Purified on Biotage™SNAP 10 g silica gel cartridge using a gradient of MeOH in CH₂Cl₂(0-20%). Pure fractions are combined and concentrated to provide titlecompound (36 mg, 0.056 mmol, 75% yield) as a beige foamy solid. ¹H NMR(400 MHz, CD₃OD) δ 8.05 (s, 1H), 8.00 (s, 1H), 7.94 (d, J=8.7 Hz, 1H),7.90 (d, J=8.6 Hz, 1H), 7.66 (dd, J=8.7, 1.6 Hz, 1H), 7.56 (dd, J=8.5,1.5 Hz, 1H), 6.03-5.97 (m, 1H), 5.32-5.25 (m, 2H), 5.22 (dd, J=8.3, 3.1Hz, 1H), 4.92 (d, J=2.1 Hz, 1H), 4.50 (dd, J=12.1, 6.7 Hz, 1H), 4.18(dd, J=12.2, 3.0 Hz, 1H), 4.06 (dd, J=3.2, 2.1 Hz, 1H), 3.99 (dd, J=9.3,3.3 Hz, 1H), 3.94-3.81 (m, 3H), 3.76 (dd, J=11.7, 5.8 Hz, 1H), 3.67 (t,J=9.6 Hz, 1H), 2.12-2.07 (m, 9H), 2.04 (s, 3H). LC-MS: m/z=645.43(M+H⁺).

Step VI, Compound 56

To a solution of[(2R,3R,4R,5R,6R)-3,4,5-triacetoxy-6-[6-[2-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]ethynyl]-2-naphthyl]tetrahydropyran-2-yl]methylacetate (34 mg, 0.053 mmol) in MeOH (1 mL) is added MeONa in MeOH (55 μLof 0.5 M, 0.028 mmol). The reaction mixture is stirred overnight thenpassed through a pre-washed SCX-2 1 g cartridge, washed with MeOH (3×1mL). Combined filtrates are concentrated to provide a glassy solid whichis redissolved in MeCN/H₂O mixture (20% MeCN) and freeze-dried toprovide title compound (24 mg, 0.046 mmol, 88% yield) as a fluffy whitesolid. ¹H NMR (400 MHz, CD₃OD) δ 7.91 (s, 1H), 7.84 (s, 1H), 7.77 (t,J=8.0 Hz, 2H), 7.61 (dd, J=8.7, 1.4 Hz, 1H), 7.40 (dd, J=8.5, 1.5 Hz,1H), 5.02 (d, J=3.9 Hz, 1H), 4.81 (d, J=2.1 Hz, 1H), 4.50-4.35 (m, 1H),3.95 (dd, J=3.2, 2.2 Hz, 1H), 3.89 (dd, J=9.3, 3.3 Hz, 1H), 3.84-3.72(m, 4H), 3.71-3.61 (m, 2H), 3.60-3.51 (m, 2H), 3.47-3.40 (m, 1H). LC-MS:m/z=477.41 (M+H⁺).

Example 38. Preparation of Compound 57 (Method D)(2R,3S,4R,5S,6R)-2-(Hydroxymethyl)-6-[2-[3-[2-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]ethynyl]phenyl]ethynyl]tetrahydropyran-3,4,5-triol

Intermediate M (108 mg, 0.57 mmol) is dissolved in DMF (2.7 mL),degassed, then added 1,3-diiodobenzene (87 mg, 0.26 mmol), followed byCuI (21 mg, 0.11 mmol) and PdCl₂(dppf). CH₂Cl₂ (22 mg, 0.027 mmol),degassed again, then Et₃N (180 μL, 1.291 mmol) is added and the reactionmixture is heated at 40° C. for 6 h. The reaction mixture isconcentrated to dryness, then purified twice by reverse-phase flashchromatography on Biotage™ SNAP C18 12+M cartridge using MeCN in H₂O,gradient of 0-30%. Pure fractions from the second purification areconcentrated to dryness, providing title compound (50 mg, 0.11 mmol, 40%yield) as a pale yellow solid. ¹H NMR (400 MHz, dmso-d6) δ 7.51-7.38 (m,4H), 4.95 (d, J=4.4 Hz, 2H), 4.76 (d, J=6.1 Hz, 2H), 4.73-4.68 (m, 4H),4.47 (t, J=6.0 Hz, 2H), 3.81-3.75 (m, 2H), 3.70-3.61 (m, 4H), 3.57-3.50(m, 2H), 3.48-3.39 (m, 2H), 3.39-3.29 (m, 6H).

LC-MS: m/z=451.49 (M+H⁺).

Example 39. Preparation of Compound 58 (Method D)(2R,3S,4R,5S,6R)-2-(Hydroxymethyl)-6-[2-[7-[2-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]ethynyl]-2-naphthyl]ethynyl]tetrahydropyran-3,4,5-triol

A microwave vial is charged with 2,7-dibromonaphthalene (36 mg, 0.13mmol) and Intermediate M (54 mg, 0.29 mmol), PdCl₂ (dppf). CH₂Cl₂ (9.2mg, 0.011 mmol) and CuI (14 mg, 0.074 mmol). DMF (1 mL) and Et₃N (88 μL,0.6295 mmol) are added, the reaction mixture is degassed and heated inmicrowave for 10 min at 120° C., then concentrated to dryness andpurified twice by reverse-phase flash chromatography on Biotage™ SNAPC18 12+M using MeCN in H₂O, gradient of 10-90% for the firstpurification, 10-60% for the second purification. Fractions from thesecond purification are concentrated to dryness and purified on prepHPLC to afford title compound (11 mg, 0.020 mmol, 16% yield) as anoff-white solid. ¹H NMR (400 MHz, DMSO-D₆) δ 8.11 (s, 2H), 7.96 (d,J=8.5 Hz, 2H), 7.54 (dd, J=8.5, 1.4 Hz, 2H), 5.01 (d, J=3.2 Hz, 2H),4.84 (d, J=4.9 Hz, 2H), 4.80-4.71 (m, 4H), 4.53 (t, J=5.6 Hz, 2H), 3.85(s, 2H), 3.71 (dd, J=18.8, 6.8 Hz, 4H), 3.65-3.55 (m, 2H), 3.52-3.20 (m,4H).

¹H NMR (400 MHz, DMSO-D₆+D₂O) δ 8.09 (s, 2H), 7.95 (d, J=8.6 Hz, 2H),7.53 (dd, J=8.5, 1.4 Hz, 2H), 4.77 (d, J=2.1 Hz, 2H), 3.86-3.81 (m, 2H),3.73 (dd, J=9.2, 3.3 Hz, 2H), 3.69 (dd, J=11.7, 1.9 Hz, 2H), 3.64-3.56(m, 2H), 3.56-3.43 (m, 2H), 3.39 (t, J=9.4 Hz, 2H). LC-MS: m/z=501.65(M+H⁺).

Preparation of Compound 59 (Method D)

(2R,3S,4R,5S,6R)-2-(Hydroxymethyl)-6-[2-[6-[2-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]ethynyl]-2-naphthyl]ethynyl]tetrahydropyran-3,4,5-triol

The title compound is obtained from 2,6-dibromonaphtalene using the sameprocedure described for compound 58 except that the reaction is carriedout at 90° C. for 16 h to afford title compound (30 mg, 0.057 mmol, 42%yield) as pale yellow solid. ¹H NMR (400 MHz, CD₃OD) δ 8.01 (s, 2H),7.86 (d, J=8.6 Hz, 2H), 7.54 (dd, J=8.5, 1.4 Hz, 2H), 4.92 (d, J=2.1 Hz,2H), 4.05 (dd, J=3.1, 2.2 Hz, 2H), 3.98 (dd, J=9.4, 3.3 Hz, 2H),3.94-3.82 (m, 4H), 3.81-3.72 (m, 2H), 3.66 (t, J=9.5 Hz, 2H). LC-MS:m/z=501.40 (M+H⁺).

Preparation of Compound 60 (Method D)

(2R,3S,4R,5S,6R)-2-(Hydroxymethyl)-6-[2-[5-[2-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]ethynyl]-1-naphthyl]ethynyl]tetrahydropyran-3,4,5-triol

The title compound is obtained from 1,5-dibromonaphtalene using the sameprocedure described for compound 58 except that the reaction is carriedout at 90° C. for 16 h to afford title compound (27 mg, 0.05 mmol, 24%yield) as an off-white fluffy/crystalline solid. ¹H NMR (400 MHz,DMSO-D6) δ 8.26 (d, J=8.5 Hz, 2H), 7.81 (d, J=6.5 Hz, 2H), 7.66 (dd,J=8.2, 7.3 Hz, 2H), 5.05 (d, J=4.4 Hz, 2H), 4.91 (d, J=2.1 Hz, 2H), 4.87(d, J=5.9 Hz, 2H), 4.82 (d, J=5.8 Hz, 2H), 4.57 (t, J=6.0 Hz, 2H),3.97-3.88 (m, 2H), 3.83-3.61 (m, 6H), 3.54-3.37 (m, 4H). LC-MS:m/z=501.54 (M+H⁺).

Preparation of Compounds 62-68

Compounds 62-64, 67 and 68 are prepared using Method A according to theprocedure described in Compound 6 using the appropriately functionalizedaryl bromide, phenol or boronic acid. Compounds 65 and 66 are preparedusing Method A according to the procedure described in Compound 139using the appropriately functionalized halogenated aryls.

LCMS Compound IUPAC mane ¹H NMR m/z (M + H+) 62(2R,2′R,3S,3′S,4R,4′R,5S,5′S,6R, (400 MHz, CD₃OD) δ 7.55 (s, 496.926′R)-6,6′-(2-fluoro-[1,1′- 4H), 7.49 (dd, J = 11.5, 4.7 Hz,biphenyl]-4,4′-diyl)bis(2- 1H), 7.34 (dd, J = 6.8, 6.3 Hz,(hydroxymethyl)tetrahydro- 2H), 5.04-4.94 (m, 2H), 2H-pyran-3,4,5-triol)4.46 (d, J = 2.3 Hz, 1H), 4.36 (d, J = 2.7 Hz, 1H), 3.90-3.79 (m, 4H),3.74 (t, J = 7.5 Hz, 2H), 3.60 (dd, J = 7.6, 2.7 Hz, 2H), 3.51 (dd, J =10.6, 6.2 Hz, 2H) 63 (2R,2′R,3S,3′S,4R,4′R,5S,5′S,6R, (400 MHz, CD₃OD) δ7.53 (d, J = 8.0 Hz, 493.35 6′R)-6,6′-(2-methyl-[1,1′- 2H), 7.38 (s,1H), biphenyl]-4,4′-diyl)bis(2- 7.31 (t, J = 8.9 Hz, 3H), 7.17 (d,(hydroxymethyl)tetrahydro- J = 7.9 Hz, 1H), 5.00 (dd, J = 15.0,2H-pyran-3,4,5-triol) 2.7 Hz, 2H), 4.48 (d, J = 3.6 Hz, 2H), 3.84 (d, J= 4.1 Hz, 4H), 3.78-3.71 (m, 2H), 3.65-3.59 (m, 2H), 3.55-3.47 (m, 2H),2.24 (s, 3H). 64 (2R,2′R,3S,3′S,4R,4′R,5S,5′S,6R, ¹H NMR (400 MHz,CD₃OD) δ 493.63 6′R)-6,6′-(3-methyl-[1,1′- 7.61 (d, J = 6.6 Hz, 2H),biphenyl]-4,4′-diyl)bis(2- 7.55-7.47 (m, 3H), 7.43 (s, 2H),(hydroxymethyl)tetrahydro- 5.13 (d, J = 6.1 Hz, 1H), 5.00 (s,2H-pyran-3,4,5-triol) 1H), 4.46 (d, J = 2.5 Hz, 1H), 4.30-4.24 (m, 1H),4.02-3.93 (m, 2H), 3.83 (s, 3H), 3.78-3.71 (m, 2H), 3.60 (dd, J = 8.0,2.7 Hz, 1H), 3.56-3.45 (m, 2H), 2.50 (s, 3H) 65(2R,2′R,3S,3′S,4R,4′R,5S,5′S,6R, (400 MHz, CD₃OD) δ N/A6′R)-6,6′-([1,1′:4′,1″- 7.75-7.68 (m, 8H), 7.59 (d, J = 8.2 Hz,terphenyl]-4,4″-diyl)bis(2- 4H), 5.05 (d, J = 3.4 Hz,(hydroxymethyl)tetrahydro- 2H), 4.50 (t, J = 3.3 Hz, 2H),2H-pyran-3,4,5-triol) 3.86 (d, J = 4.7 Hz, 4H), 3.77 (t, J = 8.1 Hz,2H), 3.64 (dd, J = 8.1, 3.1 Hz, 2H), 3.53 (dt, J = 8.3, 4.7 Hz, 2H),3.52 (dt, J = 18.3, 6.9 Hz, 2H). 66 (2R,2′R,3S,3′S,4R,4′R,5S,5′S,6R,(400 MHz, CD₃OD) δ 7.83 (t, J = 1.6 Hz, N/A 6′R)-6,6′-([1,1′:3′,1″- 1H),7.70 (d, J = 8.4 Hz, terphenyl]-4,4″-diyl)bis(2- 4H), 7.63-7.54 (m, 6H),(hydroxymethyl)tetrahydro- 7.50 (dd, J = 8.4, 6.9 Hz, 1H),2H-pyran-3,4,5-triol) 5.03 (d, J = 3.5 Hz, 2H), 4.48 (t, J = 3.3 Hz,2H), 3.84 (d, J = 4.7 Hz, 4H), 3.75 (t, J = 8.1 Hz, 2H), 3.61 (dd, J =8.1, 3.1 Hz, 2H), 3.51 (dt, J = 8.2, 4.7 Hz, 2H). 67(2R,2′R,3S,3′S,4R,4′R,5S,5′S,6R, (400 MHz, CD₃OD) δ 7.48 (s, N/A6′R)-6,6′-(2-methoxy-[1,1′- 4H), 7.30-7.21 (m, 2H),biphenyl]-4,4′-diyl)bis(2- 7.06 (d, J = 7.8 Hz, 1H),(hydroxymethyl)tetrahydro- 5.05-4.97 (m, 2H), 4.51-4.43 (m, 2H),2H-pyran-3,4,5-triol) 3.89-3.80 (m, 4H), 3.77-3.67 (m, 2H), 3.61 (dt, J= 8.1, 2.6 Hz, 2H), 3.57-3.46 (m, 2H), 3.33 (s, 3H). 68(2R,2′R,3S,3′S,4R,4′R,5S,5′S,6R, (400 MHz, CD₃OD) δ 7.50 (d, J = 8.6 Hz,N/A 6′R)-6,6′-(3,3′-dimethyl- 2H), 7.43 (d, J = 5.9 Hz,[1,1′-biphenyl]-4,4′-diyl)bis(2- 4H), 5.13 (d, J = 6.3 Hz,(hydroxymethyl)tetrahydro- 2H), 4.30-4.23 (m, 2H), 2H-pyran-3,4,5-triol)4.03-3.93 (m, 4H), 3.85-3.80 (m, 2H), 3.75 (dd, J = 11.9, 3.7 Hz, 2H),3.57-3.50 (m, 2H), 2.50 (s, 6H).Preparation of Compound 69 (Modified Method D)

(2R,3S,4R,5R,6R)-2-(hydroxymethyl)-6-[2-[4-[2-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]ethynyl]phenyl]ethynyl]tetrahydropyran-3,4,5-triol

Step I:(2R,3R,4S,5R,6R)-3,4,5-tris(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-2-ylAcetate

To a solution of(2S,3R,4S,5R,6R)-3,4,5-tribenzyloxy-6-benzyloxymethyl)tetrahydropyran-2-ol(5,920 g, 10.95 mmol) in pyridine (21.90 mL) is added acetic anhydride(2.07 mL, 21.9 mmol) dropwise. The reaction is stirred for 5 h at RTthen concentrated in vacuo. The crude mixture is diluted in CH₂Cl₂ andthe resulting solution is washed with HCl 1N and brine. The organicphase is dried over Na₂SO₄, concentrated to give a mixture of a/β of[(2R,3R,4S,5R,6R)-3,4,5-tribenzyloxy-6-(benzyloxymethyl)tetrahydropyran-2-yl]acetateas a colorless oil.

Step II:trimethyl-[2-[(2R,3S,4R,5R,6R)-3,4,5-tribenzyloxy-6-(benzyloxymethyl)tetrahydropyran-2-yl]ethynyl]silane

A mixture of[(2R,3R,4S,5R,6R)-3,4,5-tribenzyloxy-6-(benzyloxymethyl)tetrahydropyran-2-yl]acetate(1000 mg, 1.716 mmol), trimethyl-(2-tributylstannylethynyl)silane (1.329g, 1.260 mL, 3.432 mmol) and activated 4 Å molecular sieve (1 g) inCH₂Cl₂ (8.6 mL) is stirred for 15 min then TMSOTf (620 μL, 3.43 mmol) isadded dropwise at RT. The reaction is stirred is stirred for 1.5 h andEt₃N (1.00 mL, 7.18 mmol) and CH₂Cl₂ (8.6 mL) were added. The resultingmixture is filtered over celite using 10% EA/hex as eluent, the filtrateis concentrated and the residue is purified over silica gel on aBiotage™ SNAP cartridge to afford the title compound (269 mg, 25%) asyellow oil contaminated with tin byproducts.

Step III:(2R,3R,4R,5S,6R)-3,4,5-tribenzyloxy-2-(benzyloxymethyl)-6-ethynyl-tetrahydropyran

To a solution oftrimethyl-[2-[(2R,3S,4R,5R,6R)-3,4,5-tribenzyloxy-6-(benzyloxymethyl)tetrahydropyran-2-yl]ethynyl]silane(269 mg, 0.433 mmol) in MeOH (3.6 mL) and CH₂Cl₂ (722 μL) is added 1NNaOH (1.30 mL of 1 M, 1.30 mmol) for a stirring of 1 h at RT. Thereaction is quenched with 1N HCl and concentrated to remove volatiles.EtOAc is added and the organic phase is separated, dried over MgSO₄. Theresidue is dissolved in 20% EtOAc/Hex and purified over silica gel padusing 20% EA/hex as eluent. The filtrate is concentrated to give thetitle compound (83.0 mg, 35%) as a colorless oil contaminated with tinbyproducts from previous step.

Step IV:(2R,3R,4R,5S,6R)-2-ethynyl-6-(hydroxymethyl)tetrahydropyran-3,4,5-triol

To a solution of(2R,3R,4R,5S,6R)-3,4,5-tribenzyloxy-2-(benzyloxymethyl)-6-ethynyl-tetrahydropyran(76.0 mg, 0.139 mmol) in EtSH (1.4 mL) is added BF₃.OEt₂ (479 μL, 3.88mmol) and the reaction is stirred for 22 h at RT. The reaction isquenched with the addition of MeOH and basic resin to neutralize. Themixture is filtered and the filtrate is concentrated in vacuo affordingthe title compound (10 mg, 38%) as a white solid.

Preparation of(2R,3S,4R,5S,6R)-2-(hydroxymethyl)-6-[2-(4-iodophenyl)ethynyl]tetrahydropyran-3,4,5-triol

The title compound is prepared according to the procedure described forCompound 53 but using 1 eq. of 1,4-diiodobenzene in Step I: ¹H NMR (400MHz, cd3od) δ 7.66-7.59 (m, 2H), 7.15-7.08 (m, 2H), 4.75 (d, J=2.1 Hz,1H), 3.91-3.88 (m, 1H), 3.82-3.74 (m, 2H), 3.72-3.59 (m, 2H), 3.53 (t,J=9.4 Hz, 1H). LC-MS: m/z=390.94 (M+H⁺)

Step V: Compound 69

A solution of(2R,3R,4R,5S,6R)-2-ethynyl-6-(hydroxymethyl)tetrahydropyran-3,4,5-triol(10.0 mg, 0.053 mmol),(2R,3S,4R,5S,6R)-2-(hydroxymethyl)-6-[2-(4-iodophenyl)ethynyl]tetrahydropyran-3,4,5-triol(20.7 mg, 0.0531 mmol), PdCl₂(dppf). CH₂Cl₂ (8.7 mg, 0.011 mmol), CuI(2.3 mg, 0.012 mmol) in DMF (511 μL) is degased (N₂) and to this isadded triethylamine (44 μL, 0.32 mmol). The mixture is stirred in asealed tube under nitrogen atmosphere at RT for 18 h. The reaction isfiltered over 0.45 μm filter and purified by reverse phase HPLC toafford the title compound (5 mg, 20%) as a white solid. ¹H NMR (400 MHz,CD₃OD) δ 7.45-7.37 (m, 4H), 4.87-4.83 (m, 2H), 3.95 (s, 1H), 3.90-3.71(m, 5H), 3.71-3.56 (m, 4H), 3.56-3.49 (m, 1H), 3.28-3.22 (m, 1H). LC-MS:m/z=451.46 (M+H⁺)

Preparation of Compounds 70 to 76

Compound 70 is prepared according to the procedure described forCompound 53.

Compounds 71-76 are prepared according to the procedure described forCompound 59.

LCMS Compound IUPAC name ¹H-NMR m/z (M + H)⁺ 70 2-[2,5-bis[2- (400 MHz,CD3OD) δ 7.58 (d, J = 8.0 Hz, 518.38 [(2R,3S,4R,5S,6R)-3,4,5- 1H), 7.56(d, J = 1.4 Hz, 1H), trihydroxy-6- 7.45 (dd, J = 7.9, 1.5 Hz, 1H), 4.96(d, J = 2.1 Hz, (hydroxymethyl)tetrahydropyran- 1H), 4.09 (dd, J = 3.3,2.1 Hz, 2-yl]ethynyl]phenyl]-2- 1H), 4.00 (dd, J = 3.2, 2.2 Hz, 1H),methyl-propanenitrile 3.97-3.57 (m, 11H), 1.87 (s, 3H), 1.86 (s, 3H). 71(2R,3S,4R,5S,6R)-2- (400 MHz, CD3OD) δ 7.35 (d, J = 7.8 Hz, 481.41(hydroxymethyl)-6-[2-[3- 1H), 7.07 (d, J = 1.1 Hz, 1H), methoxy-4-[2-7.03 (dd, J = 7.8, 1.4 Hz, 1H), 4.87 (d, J = 2.1 Hz,[(2R,3S,4R,5S,6R)-3,4,5- 1H), 4.86 (d, J = 1.6 Hz, 1H), trihydroxy-6-4.03-3.96 (m, 3H), 3.94-3.70 (m, (hydroxymethyl)tetrahydropyran- 10H),3.69-3.58 (m, 2H). 2- yl]ethynyl]phenyl]ethynyl]tetrahydropyran-3,4,5-triol 72 (2R,3S,4R,5S,6R)-2-[2-[3- (400 MHz, CD3OD) δ 7.41 (d, J =8.0 Hz, 507.49 butyl-4-[2- 1H), 7.34 (d, J = 1.4 Hz, 1H),[(2R,3S,4R,5S,6R)-3,4,5- 7.26 (dd, J = 8.0, 1.6 Hz, 1H), 4.92 (d, J =2.0 Hz, trihydroxy-6- 1H), 4.87 (d, J = 2.1 Hz, 1H),(hydroxymethyl)tetrahydropyran- 4.05-3.95 (m, 2H), 3.93 (dd, J = 3.3, 2-1.5 Hz, 1H), 3.91 (dd, J = 3.3, 1.5 Hz, yl]ethynyl]phenyl]ethynyl]-6-1H), 3.89 (dd, J = 5.4, 1.9 Hz, 1H), (hydroxymethyl)tetrahydropyran-3.86 (dd, J = 4.9, 1.8 Hz, 1H), 3,4,5-triol 3.84-3.71 (m, 4H), 3.70-3.59(m, 2H), 2.80-2.73 (m, 2H), 1.66-1.56 (m, 2H), 1.40 (h, J = 7.3 Hz, 2H),0.96 (t, J = 7.3 Hz, 3H). 73 (2R,3S,4R,5S,6R)-2-[2-[3,5- (400 MHz,CD3OD) δ 7.20 (s, 2H), 507.49 diethyl-4-[2- 4.98 (d, J = 2.0 Hz, 1H),[(2R,3S,4R,5S,6R)-3,4,5- 4.90-4.83 (m, 1H), 4.04 (dd, J = 3.2, 2.1 Hz,trihydroxy-6- 1H), 4.00 (dd, J = 3.2, 2.1 Hz, 1H),(hydroxymethyl)tetrahydropyran- 3.94 (dd, J = 3.3, 1.6 Hz, 1H), 2- 3.92(dd, J = 3.3, 1.7 Hz, 1H), 3.89 (dd, J = 3.4,yl]ethynyl]phenyl]ethynyl]-6- 2.2 Hz, 1H), 3.86 (dd, J = 3.3, 2.2 Hz,(hydroxymethyl)tetrahydropyran- 1H), 3.84-3.78 (m, 2H), 3.76 (dd,3,4,5-triol J = 5.7, 2.2 Hz, 1H), 3.73 (dd, J = 5.7, 2.3 Hz, 1H),3.70-3.59 (m, 2H), 2.80 (q, J = 7.6 Hz, 4H), 1.24 (t, J = 7.5 Hz, 6H).74 (2R,3S,4R,5S,6R)-2- (400 MHz, CD3OD) δ 7.46-7.38 (m, 509.49(hydroxymethyl)-6-[2-[3-(2- 2H), 7.30 (dd, J = 8.0, 1.6 Hz, 1H),methoxyethyl)-4-[2- 4.92 (d, J = 2.0 Hz, 1H), 4.87 (d, J = 2.2 Hz,[(2R,3S,4R,5S,6R)-3,4,5- 1H), 4.06-3.98 (m, 2H), trihydroxy-6- 3.96-3.84(m, 4H), 3.84-3.70 (m, 4H), (hydroxymethyl)tetrahydropyran- 3.69-3.57(m, 4H), 3.35 (s, 3H), 2- 3.03 (t, J = 6.8 Hz, 2H).yl]ethynyl]phenyl]ethynyl]tetrahydropyran- 3,4,5-triol 75(2R,3S,4R,5S,6R)-2-[2-[2,3- (400 MHz, CD3OD) δ 7.49 (s, 2H), 520.34dichloro-4-[2- 4.93 (d, J = 1.9 Hz, 2H), [(2R,3S,4R,5S,6R)-3,4,5-4.07-4.01 (m, 2H), 3.96 (dd, J = 9.3, 3.2 Hz, trihydroxy-6- 2H),3.92-3.79 (m, 4H), 3.74 (dd, J = 11.5, (hydroxymethyl)tetrahydropyran-5.6 Hz, 2H), 3.66 (t, J = 9.5 Hz, 2- 2H). yl]ethynyl]phenyl]ethynyl]-6-(hydroxymethyl)tetrahydropyran- 3,4,5-triol 76(2R,3S,4R,5S,6R)-2-[2-[2,5- (400 MHz, CD3OD + dmso) δ 7.36 (s, 535.53diisopropyl-4-[2- 2H), 4.94 (d, J = 2.0 Hz, 2H),[(2R,3S,4R,5S,6R)-3,4,5- 4.03 (dd, J = 3.1, 2.2 Hz, 2H), trihydroxy-6-3.95-3.84 (m, 4H), 3.84-3.70 (m, 4H), 3.65 (t, J = 9.4 Hz,(hydroxymethyl)tetrahydropyran- 2H), 3.43-3.26 (m, 2H), 2- 1.27 (dd, J =6.9, 1.9 Hz, 12H). yl]ethynyl]phenyl]ethynyl]-6-(hydroxymethyl)tetrahydropyran- 3,4,5-triolPreparation of Compound 77 (Method D)

(2R,3S,4R,5S,6R)-2-(hydroxymethyl)-6-[2-[3-phenyl-4-[2-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]ethynyl]phenyl]ethynyl]tetrahydropyran-3,4,5-triol

[3-phenyl-4-(trifluoromethylsulfonyloxy)phenyl]trifluoromethanesulfonate(prepared from triflation of 2-phenyl hydroquinone) (54 mg, 0.12 mmol),CuI (9.0 mg, 0.048 mmol) and Pd(dppf)Cl₂— CH₂Cl₂ (9.0 mg, 0.012 mmol)are loaded in a reaction tube. A degassed solution of Intermediate M(0.5 mL of 0.53 M, 0.27 mmol) in DMF is added, followed by DIEA (105 μL,0.60 mmol). The tube is sealed and shaken at 90° C. overnight using areaction block. The reaction mixture is filtered, rinsing with DMF. Theresulting filtrate is purified by reverse phase HPLC and the fractionscontaining the desired material are combined and freeze-dried to providethe title compound (28 mg, 44% yield) as a white fluffy solid.

¹H NMR (400 MHz, CD₃OD) δ 7.56 (d, J=8.0 Hz, 1H), 7.53-7.35 (m, 7H),4.88 (d, J=2.1 Hz, 1H), 4.74 (d, J=2.0 Hz, 1H), 4.01 (dd, J=3.2, 2.1 Hz,1H), 3.93 (dd, J=9.4, 3.3 Hz, 1H), 3.88 (dd, J=11.5, 2.1 Hz, 1H),3.84-3.50 (m, 9H). ESI-MS m/z calc. 526.18, found 527.44 (M+1)⁺.

Preparation of Compounds 78 to 102

Compounds 78-84 and 86-89 are prepared according to the proceduredescribed for Compound 59. Compound 85 is prepared according to theprocedure described for Compound 171. Compounds 100-102 are preparedaccording to the procedure described for Compound 103.

LCMS Compound IUPAC name ¹H-NMR m/z (M + H)⁺ 78(2R,3S,4R,5S,6R)-2-[2-[2,3- (400 MHz, CD3OD) δ 487.43difluoro-4-[2-[(2R,3S,4R,5S,6R)- 7.38-7.24 (m, 2H), 4.92 (d, J = 2.0 Hz,3,4,5-trihydroxy-6- 2H), 4.09-3.98 (m, 2H),(hydroxymethyl)tetrahydropyran-2- 3.96-3.83 (m, 4H),yl]ethynyl]phenyl]ethynyl]-6- 3.82-3.71 (m, 4H), 3.65 (t, J = 9.4 Hz,(hydroxymethyl)tetrahydropyran- 2H). 3,4,5-triol 79 (2R,3S,4R,5S,6R)-2-(400 MHz, CD3OD) δ 7.54 (d, 495.51 (hydroxymethyl)-6-[2-[3- J = 0.9 Hz,1H), 7.46 (d, J = 7.9 Hz, (methoxymethyl)-4-[2- 1H), 7.38 (dd, J = 8.0,1.6 Hz, [(2R,3S,4R,5S,6R)-3,4,5-trihydroxy- 1H), 4.92 (d, J = 2.0 Hz,6-(hydroxymethyl)tetrahydropyran- 1H), 4.88 (d, J = 2.0 Hz, 1H),2-yl]ethynyl]phenyl]ethynyl]tetrahydropyran- 4.58 (s, 2H), 4.05-3.99 (m,3,4,5-triol 2H), 3.97-3.85 (m, 4H), 3.84-3.70 (m, 4H), 3.68-3.60 (m,2H), 3.45 (s, 3H). 80 (2R,3S,4R,5S,6R)-2-[2-[3- (400 MHz, CD3OD) δ557.51 benzyloxy-4-[2-[(2R,3S,4R,5S,6R)- 7.52-7.45 (m, 2H), 7.44-7.36(m, 3,4,5-trihydroxy-6- 3H), 7.35-7.25 (m, 1H),(hydroxymethyl)tetrahydropyran-2- 7.14 (d, J = 1.3 Hz, 1H), 7.04 (dd, J= 7.9, yl]ethynyl]phenyl]ethynyl]-6- 1.4 Hz, 1H), 5.18 (s, 2H),(hydroxymethyl)tetrahydropyran- 4.88 (d, J = 2.0 Hz, 1H), 3,4,5-triol4.87-4.83 (m, 1H), 4.03-3.99 (m, 2H), 3.97 (dd, J = 9.3, 3.3 Hz, 1H),3.93-3.86 (m, 2H), 3.85-3.59 (m, 7H). 81 (2R,3S,4R,5S,6R)-2-[2-[2,5-(400 MHz, CD3OD + dmso) δ 603.49 diphenyl-4-[2-[(2R,3S,4R,5S,6R)- 7.62(s, 2H), 7.62-7.57 (m, 3,4,5-trihydroxy-6- 4H), 7.54-7.47 (m, 4H),(hydroxymethyl)tetrahydropyran-2- 7.46-7.39 (m, 2H), 4.76 (d, J = 2.1Hz, yl]ethynyl]phenyl]ethynyl]-6- 2H), 3.84 (dd, J = 3.2, 2.1 Hz,(hydroxymethyl)tetrahydropyran- 2H), 3.77 (dd, J = 11.7, 1.4 Hz,3,4,5-triol 2H), 3.73-3.63 (m, 4H), 3.62-3.54 (m, 4H). 82(2R,3S,4R,5S,6R)-2-[2-[2,5-dihexyl- (400 MHz, CD3OD) δ 7.26 (s, 619.634-[2-[(2R,3S,4R,5S,6R)-3,4,5- 2H), 4.89 (d, J = 2.1 Hz, 2H),trihydroxy-6- 4.00 (dd, J = 3.2, 2.1 Hz, 2H),(hydroxymethyl)tetrahydropyran-2- 3.91 (dd, J = 9.3, 3.3 Hz, 2H),yl]ethynyl]phenyl]ethynyl]-6- 3.85 (dd, J = 11.2, 1.9 Hz, 2H),(hydroxymethyl)tetrahydropyran- 3.82-3.70 (m, 4H), 3.65 (t, J = 9.4 Hz,3,4,5-triol 2H), 2.76-2.58 (m, 4H), 1.67-1.50 (m, 4H), 1.45-1.22 (m,12H), 0.98-0.81 (m, 6H). 83 (2R,3S,4R,5S,6R)-2- (400 MHz, CD3OD) δ 7.34(d, 509.45 (hydroxymethyl)-6-[2-[3- J = 7.9 Hz, 1H), 7.06 (d, J = 1.2Hz, isopropoxy-4-[2-[(2R,3S,4R,5S,6R)- 1H), 7.00 (dd, J = 7.9, 1.4 Hz,3,4,5-trihydroxy-6- 1H), 4.88-4.84 (m, 2H),(hydroxymethyl)tetrahydropyran-2- 4.67 (hept, J = 6.2 Hz, 1H),yl]ethynyl]phenyl]ethynyl]tetrahydropyran- 4.09-3.96 (m, 3H),3,4,5-triol 3.95-3.83 (m, 4H), 3.77 (ddt, J = 17.4, 11.6, 5.8 Hz, 3H),3.65 (dt, J = 19.0, 7.5 Hz, 2H), 1.36 (d, J = 6.0 Hz, 6H). 84(2R,3S,4R,5S,6R)-2- (400 MHz, CD3OD) δ 4.99 (d, 507.41(hydroxymethyl)-6-[2-[2,3,5,6- J = 2.0 Hz, 2H), 4.05 (dd, J = 3.1,tetramethyl-4-[2-[(2R,3S,4R,5S,6R)- 2.2 Hz, 2H), 3.95 (dd, J = 9.3,3,4,5-trihydroxy-6- 3.3 Hz, 2H), 3.91-3.79 (m,(hydroxymethyl)tetrahydropyran-2- 4H), 3.74 (dd, J = 11.5, 5.6 Hz,yl]ethynyl]phenyl]ethynyl]tetrahydropyran- 2H), 3.66 (t, J = 9.4 Hz,2H), 3,4,5-triol 2.42 (s, 12H). 85 (2R,3S,4R,5S,6R)-2-[2-[2,5- (400 MHz,CD3OD) δ 7.59 (s, 511.47 bis(hydroxymethyl)-4-[2- 2H), 4.93 (d, J = 2.1Hz, 2H), [(2R,3S,4R,5S,6R)-3,4,5-trihydroxy- 4.74 (s, 4H), 4.03 (dd, J =3.2, 6-(hydroxymethyl)tetrahydropyran- 2.1 Hz, 2H), 3.93 (dd, J = 9.3,2-yl]ethynyl]phenyl]ethynyl]-6- 3.3 Hz, 2H), 3.88 (dd, J = 11.5,(hydroxymethyl)tetrahydropyran- 2.1 Hz, 2H), 3.81 (ddd, J = 9.5,3,4,5-triol 5.8, 2.0 Hz, 2H), 3.74 (dd, J = 11.5, 5.8 Hz, 2H), 3.65 (t,J = 9.5 Hz, 2H). 86 (2R,3S,4R,5S,6R)-2-[2-[2,5-diethyl- (400 MHz, CD3OD)δ 7.31 (s, 507.51 4-[2-[(2R,3S,4R,5S,6R)-3,4,5- 2H), 4.92 (d, J = 2.0Hz, 2H), trihydroxy-6- 4.02 (dd, J = 3.2, 2.1 Hz, 2H),(hydroxymethyl)tetrahydropyran-2- 3.93 (dd, J = 9.3, 3.3 Hz, 2H),yl]ethynyl]phenyl]ethynyl]-6- 3.88 (dd, J = 11.5, 2.1 Hz, 2H),(hydroxymethyl)tetrahydropyran- 3.84-3.78 (m, 2H), 3.74 (dd, J = 11.5,3,4,5-triol 5.7 Hz, 2H), 3.65 (t, J = 9.5 Hz, 2H), 2.75 (q, J = 7.6 Hz,4H), 1.23 (t, J = 7.6 Hz, 6H). 87 (2R,3S,4R,5S,6R)-2- (400 MHz, CD3OD) δ7.59 (d, 535.43 (hydroxymethyl)-6-[2-[3- J = 8.0 Hz, 1H), 7.50-7.42 (m,(trifluoromethoxy)-4-[2- 2H), 4.92 (d, J = 2.0 Hz, 1H),[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy- 4.89 (d, J = 2.1 Hz, 1H),6-(hydroxymethyl)tetrahydropyran- 4.04-3.98 (m, 2H), 3.93-3.82 (m, 2-4H), 3.81-3.59 (m, 6H). yl]ethynyl]phenyl]ethynyl]tetrahydropyran-3,4,5-triol 88 (2R,3S,4R,5S,6R)-2-[2-[2,5- (400 MHz, CD3OD) δ 7.36 (t, J= 7.4 Hz, 487.38 difluoro-4-[2-[(2R,3S,4R,5S,6R)- 2H), 4.91 (d, J = 2.1Hz, 3,4,5-trihydroxy-6- 2H), 4.01 (dd, J = 3.1, 2.2 Hz,(hydroxymethyl)tetrahydropyran-2- 2H), 3.93-3.83 (m, 4H),yl]ethynyl]phenyl]ethynyl]-6- 3.81-3.70 (m, 4H), 3.64 (t, J = 9.4 Hz,(hydroxymethyl)tetrahydropyran- 2H). 3,4,5-triol 89(2R,3S,4R,5S,6R)-2-[2-[2,5- (400 MHz, CD3OD + dmso) δ 539.53bis(methoxymethyl)-4-[2- 7.56 (s, 2H), 4.93 (d, J = 1.9 Hz,[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy- 2H), 4.58 (s, 4H),6-(hydroxymethyl)tetrahydropyran- 4.06-4.00 (m, 2H), 3.96-3.85 (m,2-yl]ethynyl]phenyl]ethynyl]-6- 4H), 3.85-3.69 (m, 4H),(hydroxymethyl)tetrahydropyran- 3.64 (t, J = 9.6 Hz, 2H), 3.45 (s, 6H).3,4,5-triol 90 (2R,3S,4R,5S,6R)-2-[2-[2-chloro-5- (400 MHz, CD3OD) δ7.52 (s, 499.43 methyl-4-[2-[(2R,3S,4R,5S,6R)- 1H), 7.44 (s, 1H), 4.92(d, J = 2.1 Hz, 3,4,5-trihydroxy-6- 1H), 4.91 (d, J = 2.0 Hz,(hydroxymethyl)tetrahydropyran-2- 1H), 4.08-4.01 (m, 2H),yl]ethynyl]phenyl]ethynyl]-6- 4.00-3.70 (m, 8H), 3.69-3.59 (m,(hydroxymethyl)tetrahydropyran- 2H), 2.40 (s, 3H). 3,4,5-triol 91(2R,3S,4R,5S,6R)-2- (400 MHz, CD3OD) δ 7.80 (s, 519.41(hydroxymethyl)-6-[2-[3- 1H), 7.74-7.63 (m, 2H), (trifluoromethyl)-4-[2-4.92 (d, J = 1.7 Hz, 1H), 4.90 (d, J = 2.0 Hz,[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy- 1H), 4.05-4.00 (m,6-(hydroxymethyl)tetrahydropyran- 2H), 3.96-3.58 (m, 10H). 2-yl]ethynyl]phenyl]ethynyl]tetrahydropyran- 3,4,5-triol 92(2R,3S,4R,5S,6R)-2-[2-[3- (400 MHz, CD3OD) δ 7.52 (d, 517.4(difluoromethoxy)-4-[2- J = 8.0 Hz, 1H), 7.33 (dd, J = 8.0[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy- 1.5 Hz, 1H), 7.31 (s, 1H),6-(hydroxymethyl)tetrahydropyran- 6.92 (t, J = 73.1 Hz, 1H),2-yl]ethynyl]phenyl]ethynyl]-6- 4.90 (d, J = 1.9 Hz, 1H), 4.88 (d, J =2.0 Hz, (hydroxymethyl)tetrahydropyran- 1H), 4.12-3.98 (m, 3,4,5-triol2H), 3.96-3.70 (m, 8H), 3.65 (dd, J = 19.5, 9.6 Hz, 2H). 93(2R,3S,4R,5S,6R)-2-[2-[3,5- (400 MHz, CD3OD) δ 487.48difluoro-4-[2-[(2R,3S,4R,5S,6R)- 7.23-7.15 (m, 2H), 4.94 (d, J = 2.0 Hz,3,4,5-trihydroxy-6- 1H), 4.88 (d, J = 2.1 Hz,(hydroxymethyl)tetrahydropyran-2- 1H), 4.08-3.97 (m, 2H),yl]ethynyl]phenyl]ethynyl]-6- 3.95-3.83 (m, 4H), 3.82-3.57 (m,(hydroxymethyl)tetrahydropyran- 6H). 3,4,5-triol 94(2R,3S,4R,5S,6R)-2-[2-[3-(2- (400 MHz, CD3OD) δ 495.33hydroxyethyl)-4-[2- 7.44-7.37 (m, 2H), 7.28 (d, J = 7.8 Hz,[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy- 1H), 4.90 (s, 2H), 3.99 (d, J = 9.0Hz, 6-(hydroxymethyl)tetrahydropyran- 2H), 3.92-3.82 (m,2-yl]ethynyl]phenyl]ethynyl]-6- 4H), 3.82-3.68 (m, 6H),(hydroxymethyl)tetrahydropyran- 3.61 (t, J = 9.4 Hz, 2H), 2.98 (t, J =6.9 Hz, 3,4,5-triol 2H). 95 (2R,3S,4R,5S,6R)-2- (400 MHz, CD3OD) δ 7.62(d, 481.41 (hydroxymethyl)-6-[2-[3- J = 0.8 Hz, 1H), 7.44 (d, J = 7.9Hz, (hydroxymethyl)-4-[2- 1H), 7.35 (dd, J = 7.9, 1.7 Hz,[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy- 1H), 4.92 (d, J = 2.1 Hz,6-(hydroxymethyl)tetrahydropyran- 1H), 4.88 (d, J = 2.1 Hz, 1H), 2- 4.74(s, 2H), 4.04-3.99 (m, yl]ethynyl]phenyl]ethynyl]tetrahydropyran- 2H),3.92 (ddd, J = 9.3, 3.3, 1.3 Hz, 3,4,5-triol 2H), 3.88 (dt, J = 11.4,1.7 Hz, 2H), 3.84-3.77 (m, 2H), 3.77-3.70 (m, 2H), 3.68-3.61 (m, 2H). 96(2R,3S,4R,5S,6R)-2-[2-[2,5- (400 MHz, CD3OD) δ 7.01 (s, 511.43dimethoxy-4-[2-[(2R,3S,4R,5S,6R)- 2H), 4.98-4.74 (m, 2H),3,4,5-trihydroxy-6- 4.08-3.96 (m, 4H), 3.91-3.79 (m,(hydroxymethyl)tetrahydropyran-2- 10H), 3.74 (dd, J = 11.9, 5.9 Hz,yl]ethynyl]phenyl]ethynyl]-6- 2H), 3.69-3.59 (m, 2H).(hydroxymethyl)tetrahydropyran- 3,4,5-triol 97(2R,3S,4R,5S,6R)-2-[2-[2,5- (400 MHz, CD3OD) δ 7.30 (s, 479.45dimethyl-4-[2-[(2R,3S,4R,5S,6R)- 2H), 4.91 (d, J = 2.0 Hz, 2H),3,4,5-trihydroxy-6- 4.02 (dd, J = 3.0, 2.2 Hz, 2H),(hydroxymethyl)tetrahydropyran-2- 3.94 (dd, J = 9.3, 3.2 Hz, 2H),yl]ethynyl]phenyl]ethynyl]-6- 3.88 (dd, J = 11.5, 2.0 Hz, 2H),(hydroxymethyl)tetrahydropyran- 3.82 (ddd, J = 9.4, 5.7, 2.0 Hz,3,4,5-triol 2H), 3.74 (dd, J = 11.5, 5.7 Hz, 2H), 3.64 (t, J = 9.5 Hz,2H), 2.37 (s, 6H). 98 (2R,3S,4R,5S,6R)-2-[2-[3-ethyl-4- (400 MHz, CD3OD)δ 7.41 (d, 479.49 [2-[(2R,3S,4R,5S,6R)-3,4,5- J = 8.0 Hz, 1H), 7.36 (d,J = 1.3 Hz, trihydroxy-6- 1H), 7.27 (dd, J = 7.9, 1.6 Hz,(hydroxymethyl)tetrahydropyran-2- 1H), 4.92 (d, J = 2.0 Hz,yl]ethynyl]phenyl]ethynyl]-6- 1H), 4.87 (d, J = 2.2 Hz, 1H),(hydroxymethyl)tetrahydropyran- 4.07-3.99 (m, 2H), 3.92 (dd, J = 9.3,3,4,5-triol 3.3 Hz, 2H), 3.90-3.85 (m, 2H), 3.84-3.77 (m, 2H), 3.77-3.70(m, 2H), 3.69-3.58 (m, 2H), 2.80 (q, J = 7.6 Hz, 2H), 1.25 (t, J = 7.6Hz, 3H). 99 (2R,3S,4R,5S,6R)-2-[2-[2,3- (400 MHz, CD3OD) δ 7.26 (s,479.4 dimethyl-4-[2-[(2R,3S,4R,5S,6R)- 2H), 4.92 (d, J = 2.1 Hz, 2H),3,4,5-trihydroxy-6- 4.02 (dd, J = 3.2, 2.1 Hz, 2H),(hydroxymethyl)tetrahydropyran-2- 3.94 (dd, J = 9.3, 3.3 Hz, 2H),yl]ethynyl]phenyl]ethynyl]-6- 3.88 (dd, J = 11.5, 2.1 Hz, 2H),(hydroxymethyl)tetrahydropyran- 3.81 (ddd, J = 9.5, 5.7, 2.0 Hz,3,4,5-triol 2H), 3.74 (dd, J = 11.5, 5.7 Hz, 2H), 3.65 (t, J = 9.5 Hz,2H), 2.43 (s, 6H). 100 (2R,3S,4R,5S,6R)-2-[2-[3,5- (400 MHz, CD3OD) δ7.19 (s, 479.4 dimethyl-4-[2-[(2R,3S,4R,5S,6R)- 2H), 4.98 (d, J = 2.0Hz, 1H), 3,4,5-trihydroxy-6- 4.88-4.83 (m, 1H), 4.04 (dd, J = 3.2,(hydroxymethyl)tetrahydropyran-2- 2.1 Hz, 1H), 3.99 (dd, J = 3.2,yl]ethynyl]phenyl]ethynyl]-6- 2.1 Hz, 1H), 3.98-3.85 (m,(hydroxymethyl)tetrahydropyran- 4H), 3.84-3.77 (m, 2H), 3,4,5-triol3.77-3.70 (m, 2H), 3.67 (d, J = 9.3 Hz, 1H), 3.62 (d, J = 9.4 Hz, 1H),2.41 (s, 6H). 101 2,5-bis[2-[(2R,3S,4R,5S,6R)-3,4,5- (400 MHz, CD3OD) δ7.91 (d, 476.32 trihydroxy-6- J = 1.4 Hz, 1H), 7.74 (dd, J = 8.2,(hydroxymethyl)tetrahydropyran-2- 1.7 Hz, 1H), 7.66 (d, J = 8.2 Hz,yl]ethynyl]benzonitrile 1H), 4.97 (d, J = 2.1 Hz, 1H), 4.89 (d, J = 2.1Hz, 1H), 4.07 (dd, J = 3.3, 2.1 Hz, 1H), 4.01 (dd, J = 3.2, 2.2 Hz, 1H),3.97 (dd, J = 9.4, 3.3 Hz, 1H), 3.92-3.59 (m, 9H). 102(2R,3S,4R,5S,6R)-2-[2-[3-chloro-4- (400 MHz, CD3OD) δ 7.59 (d, 485.33[2-[(2R,3S,4R,5S,6R)-3,4,5- J = 1.5 Hz, 1H), 7.53 (d, J = 8.0 Hz,trihydroxy-6- 1H), 7.39 (dd, J = 8.0, 1.6 Hz,(hydroxymethyl)tetrahydropyran-2- 1H), 4.92 (d, J = 2.1 Hz,yl]ethynyl]phenyl]ethynyl]-6- 1H), 4.88 (d, J = 2.1 Hz, 1H),(hydroxymethyl)tetrahydropyran- 4.03 (dd, J = 3.2, 2.1 Hz, 1H),3,4,5-triol 4.02-3.94 (m, 2H), 3.93-3.70 (m, 7H), 3.69-3.59 (m, 2H).Preparation of Compound 103 (Method C)

(2R,3S,4R,5S,6R)-2-(hydroxymethyl)-6-[2-[3-isopropyl-4-[2-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]ethynyl]phenyl]ethynyl]tetrahydropyran-3,4,5-triol

Step I:[(2R,3R,4R,5R,6R)-6-[2-[4-[2-[(2R,3R,4R,5R,6R)-6-(hydroxymethyl)-3,4,5-tris(triisopropylsilyloxy)tetrahydropyran-2-yl]ethynyl]-3-isopropyl-phenyl]ethynyl]-3,4,5-tris(triisopropylsilyloxy)tetrahydropyran-2-yl]

A glass vial is loaded with 1,4-dibromo-2-isopropyl-benzene (60 mg, 0.22mmol), iodocopper (12 mg, 0.061 mmol) and Pd(dppf)Cl₂— CH₂Cl₂ (20 mg,0.028 mmol), capped and degassed (vacuum-nitrogen flush, 3 times). Adegassed DMF solution of Intermediate L (1.6 mL of 0.26M, 0.41 mmol),and degassed DMF (0.9 mL) are added, followed by a degassed DBU and H₂Omixture (8:3 v/v, 250 μL). The vial is transferred to a preheated oilbath (90° C.) and stirred overnight. The reaction mixture is cooled toRT and partitioned between saturated NH₄Cl solution (5 mL) and EtOAc (10mL). The layers are separated and the aqueous layer is back extractedwith EtOAc (2×2 mL). The combined organic extracts are dried overNa₂SO₄, filtered and concentrated. The residue is purified by flashchromatography on a Biotage™ SNAP 25 g silica cartridge, using agradient of EtOAc in Hex, 0-20% as eluent to provide the title compound(90 mg, 29% yield) as a yellowish gum.

Step II: Compound 103

A solution of[(2R,3R,4R,5R,6R)-6-[2-[4-[2-[(2R,3R,4R,5R,6R)-6-(hydroxymethyl)-3,4,5-tris(triisopropylsilyloxy)tetrahydropyran-2-yl]ethynyl]-3-isopropyl-phenyl]ethynyl]-3,4,5-tris(triisopropylsilyloxy)tetrahydropyran-2-yl]methanol(90 mg, 0.066 mmol) from Step I in a mixture of THF (8104), TFA (405 μL)and H₂O (405 μL) is heated to reflux (80° C.) for 48 h and concentrated.The residue is purified by reverse-phase flash chromatography on aBiotage™ SNAP C18-12 g cartridge, using a gradient of MeCN in H₂O, 0-70%as eluent. The fractions are concentrated and freeze-dried, providingthe title compound (13 mg, 40% yield) as a fluffy white solid. ¹H NMR(400 MHz, CD₃OD) δ 7.41 (d, J=8.0 Hz, 1H), 7.39 (d, J=1.5 Hz, 1H), 7.26(dd, J=7.9, 1.6 Hz, 1H), 4.92 (d, J=2.1 Hz, 1H), 4.88 (d, J=2.1 Hz, 1H),4.04-3.98 (m, 2H), 3.95-3.88 (m, 3H), 3.86 (dd, J=3.3, 2.1 Hz, 1H),3.84-3.71 (m, 4H), 3.69-3.58 (m, 2H), 3.41 (p, J=6.9 Hz, 1H), 1.27 (d,J=6.9 Hz, 6H). ESI-MS m/z calc. 492.19955, found (M+1)+493.36.

Preparation of Compounds 104 to 109

Compounds 104 and 105 are prepared according to the procedure describedfor Compound 103.

Compounds 106, 108 and 109 are prepared according to the proceduredescribed for Compound 59. Compound 107 is prepared according to theprocedure described for Compound 110.

LCMS Compound IUPAC name ¹H-NMR m/z (M + H)⁺ 104 (2R,3S,4R,5S,6R)-2-(400 MHz, CD3OD) δ 7.40 (d, J = 8.0 Hz, 465.35 (hydroxymethyl)-6-[2-[3-1H), 7.37 (s, 1H), 7.26 (dd, J = 7.9, methyl-4-[2- 1.2 Hz, 1H), 4.92 (d,J = 2.1 Hz, 1H), [(2R,3S,4R,5S,6R)-3,4,5- 4.89-4.83 (m, 1H), 4.02 (dd, J= 3.2, trihydroxy-6- 2.1 Hz, 1H), 4.00 (dd, J = 3.2, 2.1 Hz,(hydroxymethyl)tetrahydropyran- 1H), 3.95 (d, J = 3.3 Hz, 1H), 3.92 (dd,2- J = 3.3, 1.4 Hz, 1H), 3.91-3.88 (m,yl]ethynyl]phenyl]ethynyl]tetrahydropyran- 1H), 3.87-3.85 (m, 1H),3,4,5-triol 3.84-3.77 (m, 2H), 3.75 (d, J = 5.5 Hz, 1H), 3.72 (d, J =5.7 Hz, 1H), 3.68-3.60 (m, 2H), 2.42 (s, 3H). 105(2R,3S,4R,5S,6R)-2-[2-[3- (400 MHz, CD3OD) δ 7.52-7.42 (m, 469.41fluoro-4-[2-[(2R,3S,4R,5S,6R)- 1H), 7.28 (dd, J = 5.9, 1.1 Hz, 1H),3,4,5-trihydroxy-6- 7.26 (dd, J = 4.1, 1.5 Hz, 1H), 4.89 (d,(hydroxymethyl)tetrahydropyran- J = 2.1 Hz, 1H), 4.85 (d, J = 2.1 Hz, 2-1H), 4.05-3.95 (m, 2H), yl]ethynyl]phenyl]ethynyl]-6- 3.93-3.82 (m, 4H),3.81-3.67 (m, 4H), (hydroxymethyl)tetrahydropyra- 3.66-3.55 (m, 2H).3,4,5-triol 106 (2R,3S,4R,5S,6R)-2- (400 MHz, CD3OD) δ 8.19 (s, 2H),541.36 (hydroxymethyl)-6-[2-[8-[2- 7.60 (s, 4H), 4.79 (s, 2H), 4.03 (d,J = 2.1 Hz, [(2R,3S,4R,5S,6R)-3,4,5- 2H), 3.98 (dd, J = 9.3, 3.2 Hz,trihydroxy-6- 2H), 3.91-3.82 (m, 4H), (hydroxymethyl)tetrahydropyran-3.78-3.71 (m, 2H), 3.68-3.61 (m, 2H). 2-yl]ethynyl]dibenzofuran-2-yl]ethynyl]tetrahydropyran- 3,4,5-triol 107 (2R,3S,4R,5S,6R)-2-[2-[9-(400 MHz, CD3OD) δ 8.23 (s, 2H), 568.4 ethyl-6-[2-[(2R,3S,4R,5S,6R)-7.53 (dd, J = 20.0, 8.1 Hz, 4H), 4.79 (s, 3,4,5-trihydroxy-6- 2H), 4.43(d, J = 8.1 Hz, 2H), (hydroxymethyl)tetrahydropyran- 4.06-3.99 (m, 4H),3.88 (d, J = 9.4 Hz, 4H), 2-yl]ethynyl]carbazol-3- 3.75 (dd, J = 11.7,5.8 Hz, 2H), yl]ethynyl]-6- 3.70-3.61 (m, 2H), 1.39 (t, J = 7.2 Hz, 3H).(hydroxymethyl)tetrahydropyran- 3,4,5-triol 108 (2R,3S,4R,5S,6R)-2- (400MHz, CD3OD) δ 7.50-7.44 (m, 540.38 (hydroxymethyl)-6-[2-[6-[2- 3H),7.41-7.35 (m, 3H), 4.85 (s, 2H), [(2R,3S,4R,5S,6R)-3,4,5- 3.98 (s, 2H),3.88 (dd, J = 19.1, 9.3 Hz, trihydroxy-6- 4H), 3.81-3.76 (m, 2H), 3.71(dd, J = 12.3, (hydroxymethyl)tetrahydropyran- 5.0 Hz, 2H), 3.66-3.57(m, 2H). 2-yl]ethynyl]-9H-carbazol-3- yl]ethynyl]tetrahydropyran-3,4,5-triol 109 (2R,3S,4R,5S,6R)-2- (400 MHz, CD3OD) δ 8.39 (s, 2H),557.31 (hydroxymethyl)-6-[2-[7-[2- 7.90 (d, J = 8.3 Hz, 2H), 7.57 (d, J= 8.3 Hz, [(2R,3S,4R,5S,6R)-3,4,5- 2H), 4.79 (s, 2H), 4.04 (dd, J = 3.0,trihydroxy-6- 2.2 Hz, 2H), 3.99 (dd, J = 9.3, 3.4 Hz,(hydroxymethyl)tetrahydropyran- 2H), 3.92-3.85 (m, 4H), 3.75 (dd, 2- J =11.5, 5.5 Hz, 2H), 3.69-3.62 (m, yl]ethynyl]dibenzothiophen-3- 2H).yl]ethynyl]tetrahydropyran- 3,4,5-triolPreparation of Compound 110 (Method D)

(2R,3S,4R,5S,6R)-2-(hydroxymethyl)-6-[2-[9-propyl-7-[2-[2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]ethynyl]carbazol-2-yl]ethynyl]tetrahydropyran-3,4,5-triol

A mixture of Intermediate M (500 μL of 0.53 M in DMF, 0.265 mmol), AG15(43 mg, 0.1171 mmol), Pd(dppf)Cl₂— CH₂Cl₂ (15 mg, 0.0184 mmol), CuI (15mg, 0.079 mmol) in DMF (200 μL) is degased (N₂). To the resultingmixture is added N-ethyl-N-isopropyl-propan-2-amine (180 μL, 1.03 mmol).The final mixture is stirred in a sealed tube under nitrogen atmosphereat 100° C. for 18h. The reaction is filtered over 0.45 μM filter andconcentrated in vacuo. The residue is purified by reverse phase HPLC toafford the title compound (7.9 mg, 10%) as a grey solid. ¹H NMR (400MHz, CD₃OD) δ 8.05 (d, J=7.9 Hz, 2H), 7.63 (s, 2H), 7.29 (d, J=7.9 Hz,2H), 4.90 (s, 2H), 4.33 (t, J=7.1 Hz, 2H), 4.04 (s, 2H), 4.03-3.96 (m,2H), 3.88 (t, J=8.0 Hz, 4H), 3.75 (dd, J=12.1, 5.5 Hz, 2H), 3.65 (t,J=9.3 Hz, 2H), 1.87 (dd, J=14.5, 7.2 Hz, 2H), 0.93 (t, J=7.1 Hz, 3H).ESI-MS m/z calc. 581.61. found 582.44 (M+1)⁺.

Preparation of Compounds 111 and 112

Compounds 111 and 112 are prepared according to the procedure describedfor Compound 110 using the appropriate Intermediates.

LCMS Compound IUPAC name ¹H-NMR m/z (M + H)⁺ 111(2R,3S,4R,5S,6R)-2-[2-[9-ethyl- (400 MHz, CD3OD) δ 8.05 (d, J = 8.1 Hz,568.53 7-[2-[(2R,3S,4R,5S,6R)-3,4,5- 2H), 7.64 (s, 2H), 7.29 (d, J = 8.3Hz, trihydroxy-6- 2H), 4.78 (s, 2H), 4.06-4.02 (m,(hydroxymethyl)tetrahydropyran- 2H), 4.00 (dd, J = 9.5, 3.4 Hz, 2H),2-yl]ethynyl]carbazol-2- 3.92-3.85 (m, 6H), 3.75 (dd, J = 12.1,yl]ethynyl]-6- 6.5 Hz, 2H), 3.69-3.62 (m,(hydroxymethyl)tetrahydropyran- 2H), 1.38 (t, J = 7.1 Hz, 3H).3,4,5-triol 112 (2R,3S,4R,5S,6R)-2- (400 MHz, CD3OD) δ 8.05 (d, J = 8.1Hz, 554.39 (hydroxymethyl)-6-[2-[9- 2H), 7.64 (s, 2H), 7.30 (d, J = 8.5Hz, methyl-7-[2-[(2R,3S,4R,5S,6R)- 2H), 4.78 (s, 2H), 4.04 (d, J = 2.9Hz, 3,4,5-trihydroxy-6- 2H), 4.03-3.97 (m, 2H),(hydroxymethyl)tetrahydropyran- 3.93-3.84 (m, 7H), 3.79-3.74 (m, 2H),2-yl]ethynyl]carbazol-2- 3.65 (t, J = 9.5 Hz, 2H).yl]ethynyl]tetrahydropyran- 3,4,5-triol

Preparation of Compound 113 (Method D)

(2R,3S,4R,5S,6R)-2-(hydroxymethyl)-6-[2-[9-hydroxy-9-pentyl-7-[2-[2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]ethynyl]fluoren-2-yl]ethynyl]tetrahydropyran-3,4,5-triol

Step I 2,7-dibromo-9-pentyl-fluoren-9-ol

A solution of 2,7-dibromofluoren-9-one (200 mg, 0.592 mmol) in THF (2mL) is added to a solution of n-pentyl magnesium bromide 2.0M in Et₂O(355 μL of 2.0 M, 0.710 mmol) diluted with Et₂O (4 mL) at 0° C. Thereaction is stirred 18h at RT and quenched with 1N Na₂CO₃. The organicphase is separated, washed with brine, dried over MgSO₄, filtered andconcentrated. The residue is purified on Biotage™ SNAP silica gelcartridge (0-8% EA/hex) to give 2,7-dibromo-9-pentyl-fluoren-9-ol (150mg, 62%) as a white solid.

Step II: Compound 113

A solution of Intermediate M (500 μL of 0.53 M in DMF, 0.265 mmol),2,7-dibromo-9-pentyl-fluoren-9-ol (50.0 mg, 0.122 mmol), Pd(dppf)Cl₂—CH₂Cl₂ (15 mg, 0.018 mmol), CuI (15 mg, 0.07876 mmol) in DMF (200 μL) isdegased (N₂). To the resulting mixture is addedN-ethyl-N-isopropyl-propan-2-amine (180 μL, 1.03 mmol). The finalmixture is stirred in a sealed tube under nitrogen atmosphere at 100° C.for 18h. The reaction is filtered over 0.45 μM filter and concentratedin vacuo. The residue is purified by reverse phase HPLC to afford thetitle compound (9.4 mg, 11%) as a white solid. ¹H NMR (400 MHz, CD₃OD) δ7.68 (d, J=7.8 Hz, 2H), 7.55 (s, 2H), 7.47 (d, J=7.8 Hz, 2H), 4.80 (s,2H), 4.01 (s, 2H), 3.98-3.92 (m, 2H), 3.91-3.80 (m, 4H), 3.73 (dd,J=11.3, 5.6 Hz, 2H), 3.63 (t, J=9.6 Hz, 2H), 2.15-2.07 (m, 2H),1.16-1.07 (m, 4H), 0.79-0.64 (m, 5H). ESI-MS m/z calc. 624.68, found625.47 (M+1)⁺.

Preparation of Compounds 114 to 119

Compounds 114 and 115 are prepared according to the procedure describedfor Compound 113 using the appropriate Intermediates. Compounds 116-119are prepared according to the procedure described for Compound 59 usingcommercially available starting material.

LCMS Compound IUPAC name ¹H-NMR m/z (M + H)⁺ 114(2R,3S,4R,5S,6R)-2-[2-[9- (400 MHz, CD3OD) δ 7.67 (d, J = 7.9 Hz, 595.5cyclopropyl-9-hydroxy-7-[2- 2H), 7.59 (s, 2H), 7.47 (d, J = 8.0 Hz,[(2R,3S,4R,5S,6R)-3,4,5- 2H), 4.80 (s, 2H), 4.01 (t, trihydroxy-6- J =2.7 Hz, 2H), 3.97-3.92 (m, (hydroxymethyl)tetrahydropyran- 2H), 3.87(dd, J = 11.7, 2.1 Hz, 2H), 2-yl]ethynyl]fluoren-2-yl]ethynyl]-3.84-3.80 (m, 2H), 3.73 (dd, J = 11.5, 6- 5.6 Hz, 2H), 3.63 (t, J = 9.5Hz, (hydroxymethyl)tetrahydropyran- 2H), 1.16-1.08 (m, 1H), 3,4,5-triol0.58 (d, J = 4.4 Hz, 2H), 0.44 (d, J = 9.3 Hz, 2H). 115(2R,3S,4R,5S,6R)-2- (400 MHz, CD3OD) δ 7.68 (d, J = 7.8 Hz, 569.6(hydroxymethyl)-6-[2-[9-hydroxy- 1H), 7.61 (s, 1H), 7.46 (dd,9-methyl-7-[2-[(2R,3S,4R,5S,6R)- J = 7.9, 1.4 Hz, 1H), 4.79 (s, 2H),3,4,5-trihydroxy-6- 4.04-3.99 (m, 2H), 3.94 (dd, J = 9.4,(hydroxymethyl)tetrahydropyran- 3.2 Hz, 2H), 3.90-3.80 (m,2-yl]ethynyl]fluoren-2- 4H), 3.73 (dd, J = 11.5, 5.6 Hz, 2H),yl]ethynyl]tetrahydropyran-3,4,5- 3.63 (t, J = 9.4 Hz, 2H), 1.65 (s,triol 3H). 116 (2R,3S,4R,5S,6R)-2-[2-[9,9- (400 MHz, CD3OD) δ 575.42difluoro-7-[2-[(2R,3S,4R,5S,6R)- 7.76-7.70 (m, 4H), 7.64 (d, J = 7.8 Hz,2H), 3,4,5-trihydroxy-6- 4.78 (d, J = 3.4 Hz, 2H),(hydroxymethyl)tetrahydropyran- 4.03-4.00 (m, 2H), 3.90 (ddd, J = 13.8,10.4, 2-yl]ethynyl]fluoren-2-yl]ethynyl]- 2.8 Hz, 4H), 3.84-3.79 (m,2H), 6- 3.73 (dd, J = 11.6, 5.8 Hz, 2H), (hydroxymethyl)tetrahydropyran-3.62 (t, J = 9.4 Hz, 2H). 3,4,5-triol 117 (2R,3S,4R,5S,6R)-2- (400 MHz,CD3OD) δ 7.76 (d, J = 8.1 Hz, 552.57 (hydroxymethyl)-6-[2-[7-[2- 2H),7.40-7.34 (m, 4H), [(2R,3S,4R,5S,6R)-3,4,5- 4.79 (s, 2H), 4.00 (dd, J =3.3, 2.1 Hz, trihydroxy-6- 2H), 3.93 (dd, J = 9.2, 3.3 Hz,(hydroxymethyl)tetrahydropyran- 2H), 3.87 (dd, J = 11.6, 2.0 Hz, 2H),2-yl]ethynyl]-9,10- 3.81 (dd, J = 8.8, 1.8 Hz, 2H),dihydrophenanthren-2- 3.73 (dd, J = 11.1, 5.2 Hz, 2H), 3.64 (d, J = 9.4Hz, yl]ethynyl]tetrahydropyran-3,4,5- 2H), 2.83 (s, 4H). triol 118(2R,3S,4R,5S,6R)-2- (400 MHz, CD3OD) δ 8.02 (d, J = 8.1 Hz, 540.42(hydroxymethyl)-6-[2-[7-[2- 2H), 7.54 (s, 2H), 7.26 (dd,[(2R,3S,4R,5S,6R)-3,4,5- J = 8.2, 1.2 Hz, 2H), 4.78 (s, 2H),trihydroxy-6- 4.03 (dd, J = 3.1, 2.3 Hz, 2H),(hydroxymethyl)tetrahydropyran- 3.98 (dd, J = 9.4, 3.1 Hz, 2H),2-yl]ethynyl]-9H-carbazol-2- 3.91-3.84 (m, 4H), 3.74 (dd, J = 11.5, 5.6Hz, yl]ethynyl]tetrahydropyran-3,4,5- 2H), 3.64 (t, J = 9.5 Hz, 2H).triol 119 2,7-bis[2-[(2R,3S,4R,5S,6R)- (400 MHz, CD3OD) δ 553.523,4,5-trihydroxy-6- 7.74-7.64 (m, 6H), 4.82 (s, 2H),(hydroxymethyl)tetrahydropyran- 4.02-3.99 (m, 2H), 3.94-3.88 (m, 4H),2-yl]ethynyl]fluoren-9-one 3.83-3.77 (m, 2H), 3.72 (dd, J = 11.5, 6.0Hz, 2H), 3.62 (t, J = 9.6 Hz, 2H).Preparation of Compound 120 (Method D)

(2R,3S,4R,5S,6R)-2-[2-[9,9-dimethyl-7-[2-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]ethynyl]fluoren-2-yl]ethynyl]-6-(hydroxymethyl)tetrahydropyran-3,4,5-triol

A solution of Intermediate M (1.00 g, 5.31 mmol),2,7-dibromo-9,9-dimethyl-fluorene (870 mg, 2.47 mmol),Pd(dppf)Cl2-CH₂Cl₂ (141 mg, 0.1727 mmol), CuI (141 mg, 0.7404 mmol) isdegased (N2) and to the resulting mixture is addedN-ethyl-N-isopropyl-propan-2-amine (3.73 mL, 21.4 mmol). The finalmixture is stirred in a sealed tube under nitrogen atmosphere at 100° C.for 1 h. The reaction is poured in a round bottom flask containing Ac₂O(25.0 mL, 265 mmol) and pyridine (25 mL, 309 mmol) and the resultingmixture is stirred over 18 h at RT. The reaction is then concentrated invacuo, the residue is dissolved in EA (100 mL) and washed with NH₄Cl(3×125 mL). The organic phase is dried over MgSO₄, filtered, andconcentrated. The resulting mixture is purified on Biotage™ SNAP 100 gsilica gel cartridge using EtOAc (20 to 80%) to afford theper-acetylated 120 (1.36 g, 61%) as a pale yellow foamy solid. ¹H NMR(400 MHz, CDCl₃) δ 7.70 (d, J=7.9 Hz, 2H), 7.57 (s, 2H), 7.52 (dd,J=7.9, 1.3 Hz, 2H), 5.60 (dd, J=10.0, 3.4 Hz, 2H), 5.48 (dd, J=3.2, 2.1Hz, 2H), 5.36 (t, J=10.0 Hz, 2H), 5.05 (d, J=2.0 Hz, 2H), 4.37 (dd,J=12.2, 4.8 Hz, 2H), 4.31 (ddd, J=10.0, 4.7, 2.0 Hz, 2H), 4.18 (dd,J=12.2, 2.0 Hz, 2H), 2.22 (s, 6H), 2.14 (s, 6H), 2.07 (s, 6H), 2.04 (s,6H), 1.52 (s, 6H). ESI-MS m/z calc. 902.29974. found 903.73 (M+1)⁺.

To the per-acetylated Compound 120 namely[(2R,3R,4R,5R,6R)-3,4,5-triacetoxy-6-[2-[9,9-dimethyl-7-[2-[(2R,3R,4R,5R,6R)-3,4,5-triacetoxy-6-(acetoxymethyl)tetrahydropyran-2-yl]ethynyl]fluoren-2-yl]ethynyl]tetrahydropyran-2-yl]methylacetate (1.36 g, 1.506 mmol) dissolved in MeOH (27 mL) (clear solution)is added MeONa in MeOH (3.00 mL of 0.5 M, 1.50 mmol). After 1h, thereaction mixture is treated with prewashed Dowex 50WX4 (2.9 g used),filtered and concentrated. To the residual brown is added 15 mL MeOH,the suspension is sonicated then stirred at 40° C. under N₂ for 1.5h.The mixture is cooled down to RT, placed in an ice bath and filtered toafford the title compound as a light beige solid (659 mg, 76% from theper-acetylated intermediate). ¹H NMR (400 MHz, CD₃OD) δ 7.74 (d, J=7.8Hz, 2H), 7.57 (s, 2H), 7.43 (d, J=7.9 Hz, 2H), 4.87 (s, 2H), 4.02 (s,2H), 3.96 (dd, J=9.2, 3.1 Hz, 2H), 3.90-3.81 (m, 4H), 3.74 (dd, J=11.2,5.2 Hz, 2H), 3.64 (t, J=9.2 Hz, 2H), 1.46 (s, 6H). [1], 1H NMR (400 MHz,cd3od) 7.74 (d, J=7.9 Hz, 2H), 7.57 (s, 2H), 7.44 (d, J=7.9 Hz, 2H),4.92 (s, 2H), 4.03-4.00 (m, 2H), 3.96 (dd, J=9.4, 3.0 Hz, 2H), 3.91-3.81(m, 4H), 3.74 (dd, J=11.2, 5.3 Hz, 2H), 3.63 (t, J=9.2 Hz, 2H), 1.46 (s,6H). ESI-MS m/z calc. 566.60. found 567.42 (M+H)⁺.

Preparation of Compounds 121 to 138

Compounds 121-138 are prepared according to the procedure described forCompound 59 using commercially available starting material.

LCMS Compound IUPAC name ¹H-NMR m/z (M + H)⁺ 121 (2R,3S,4R,5S,6R)-2-(400 MHz, CD3OD + DMSO) δ 555.55 (hydroxymethyl)-6-[2-[2-methyl- 7.55(s, 2H), 7.50 (d, J = 8.0 Hz, 2H), 4-[3-methyl-4-[2- 7.46 (dd, J = 8.1,1.6 Hz, 2H), [(2R,3S,4R,5S,6R)-3,4,5- 4.94 (d, J = 2.1 Hz, 2H), 4.04(dd, J = 3.2, trihydroxy-6- 2.1 Hz, 2H), 3.98 (dd, J = 9.4,(hydroxymethyl)tetrahydropyran- 3.3 Hz, 2H), 3.92-3.81 (m, 4H), 2- 3.75(dd, J = 11.3, 5.5 Hz, 2H),yl]ethynyl]phenyl]phenyl]ethynyl]tetrahydropyran- 3.66 (t, J = 9.4 Hz,2H), 2.51 (s, 6H). 3,4,5-triol 122 (2R,3S,4R,5S,6R)-2- (400 MHz, CD3OD)δ 569.6 (hydroxymethyl)-6-[2-[4-[1- 7.42-7.33 (m, 4H), 7.25-7.16 (m,4H), methyl-1-[4-[2- 4.85 (d, J = 2.0 Hz, 2H), 3.99 (dd, J = 3.1,[(2R,3S,4R,5S,6R)-3,4,5- 2.2 Hz, 2H), 3.93 (dd, J = 9.3, trihydroxy-6-3.3 Hz, 2H), 3.87 (dd, J = 11.5, 2.0 Hz, (hydroxymethyl)tetrahydropyran-2H), 3.80 (ddd, J = 9.5, 5.7, 2.0 Hz, 2- 2H), 3.73 (dd, J = 11.5, 5.7Hz, yl]ethynyl]phenyl]ethyl]phenyl]ethynyl]tetrahydropyran- 2H), 3.63(t, J = 9.5 Hz, 2H), 3,4,5-triol 1.66 (s, 6H). 123 (2R,3S,4R,5S,6R)-2-(400 MHz, CD3OD) δ 7.41 (s, 2H), 513.39 (hydroxymethyl)-6-[2-[5-[2- 4.89(d, J = 2.0 Hz, 2H), 3.99 (t, J = 2.6 Hz, [(2R,3S,4R,5S,6R)-3,4,5- 2H),3.90-3.82 (m, 4H), trihydroxy-6- 3.73 (dq, J = 12.3, 6.2 Hz, 4H),(hydroxymethyl)tetrahydropyran- 3.62 (dd, J = 11.3, 7.4 Hz, 2H).2-yl]ethynyl]thieno[2,3- b]thiophen-2- yl]ethynyl]tetrahydropyran-3,4,5-triol 124 (2R,3S,4R,5S,6R)-2- (400 MHz, CD3OD) δ 7.39 (s, 2H), 555.4(hydroxymethyl)-6-[2-[3-methyl- 7.34-7.29 (m, 2H), 7.03 (d, J = 7.8 Hz,4-[2-methyl-4-[2- 2H), 4.86 (d, J = 2.1 Hz, 2H),[(2R,3S,4R,5S,6R)-3,4,5- 4.00 (dd, J = 3.2, 2.2 Hz, 2H), trihydroxy-6-3.94 (dd, J = 9.3, 3.3 Hz, 2H), (hydroxymethyl)tetrahydropyran-3.89-3.80 (m, 4H), 3.73 (dd, J = 11.4, 5.5 Hz, 2- 2H), 3.63 (t, J = 9.4Hz, 2H), yl]ethynyl]phenyl]phenyl]ethynyl]tetrahydropyran- 2.00 (s, 6H).3,4,5-triol 125 (2R,3S,4R,5S,6R)-2-[2-[1,5- (400 MHz, CD3OD) δ 7.87 (d,J = 8.6 Hz, 561.46 dimethoxy-6-[2- 2H), 7.50 (d, J = 8.6 Hz,[(2R,3S,4R,5S,6R)-3,4,5- 2H), 4.97 (d, J = 2.1 Hz, 2H), trihydroxy-6-4.12 (s, 6H), 4.08 (dd, J = 3.2, 2.1 Hz, (hydroxymethyl)tetrahydropyran-2H), 4.01 (dd, J = 9.3, 3.3 Hz, 2H), 2-yl]ethynyl]-2-naphthyl]ethynyl]-3.95-3.84 (m, 4H), 3.76 (dd, J = 12.0, 6- 6.1 Hz, 2H), 3.68 (t, J = 9.5Hz, (hydroxymethyl)tetrahydropyran- 2H). 3,4,5-triol 126(2R,3S,4R,5S,6R)-2- (400 MHz, CD3OD) δ 8.33 (d, J = 8.8 Hz, 502.32(hydroxymethyl)-6-[2-[2-[2- 1H), 8.09 (s, 1H), 7.94 (d, J = 8.7 Hz,[(2R,3S,4R,5S,6R)-3,4,5- 1H), 7.80 (d, J = 8.3 Hz, trihydroxy-6- 1H),7.65 (d, J = 8.7 Hz, 1H), (hydroxymethyl)tetrahydropyran- 4.96 (s, 2H),4.11-4.02 (m, 2H), 2-yl]ethynyl]-6- 3.99-3.92 (m, 2H), 3.91-3.81 (m,4H), quinolyl]ethynyl]tetrahydropyran- 3.75 (dd, J = 11.4, 5.5 Hz, 2H),3,4,5-triol 3.69-3.61 (m, 2H). 127 (2R,3S,4R,5S,6R)-2- (400 MHz, CD3OD)δ 9.47 (s, 1H), 503.33 (hydroxymethyl)-6-[2-[2-[2- 8.25 (d, J = 1.6 Hz,1H), 8.04 (dd, J = 8.8, [(2R,3S,4R,5S,6R)-3,4,5- 1.8 Hz, 1H), 7.95 (d, J= 8.8 Hz, trihydroxy-6- 1H), 4.96 (d, J = 2.1 Hz, 1H),(hydroxymethyl)tetrahydropyran- 4.92 (d, J = 2.1 Hz, 1H), 4.09 (dd, J =3.2, 2-yl]ethynyl]quinazolin-6- 2.2 Hz, 1H), 4.04 (dd, J = 3.2,yl]ethynyl]tetrahydropyran-3,4,5- 2.2 Hz, 1H), 3.98-3.91 (m, triol 2H),3.89 (dd, J = 5.2, 2.9 Hz, 1H), 3.88-3.84 (m, 2H), 3.84-3.79 (m, 1H),3.78-3.71 (m, 2H), 3.65 (dd, J = 19.3, 9.7 Hz, 2H). 128(2R,3S,4R,5S,6R)-2- (400 MHz, CD3OD) δ 7.18 (dd, J = 10.4, 539.39(hydroxymethyl)-6-[2-[5-[5-[2- 3.7 Hz, 4H), 4.88 (s, 2H),[(2R,3S,4R,5S,6R)-3,4,5- 3.98 (d, J = 2.0 Hz, 2H), 3.86 (dd, J = 9.4,trihydroxy-6- 3.2 Hz, 4H), 3.77-3.68 (m, (hydroxymethyl)tetrahydropyran-4H), 3.62 (t, J = 9.4 Hz, 2H). 2-yl]ethynyl]-2-thienyl]-2-thienyl]ethynyl]tetrahydropyran- 3,4,5-triol 129 (2R,3S,4R,5S,6R)-2-(400 MHz, CD3OD) δ 8.20 (s, 1H), 442.029 (hydroxymethyl)-6-[2-[2-[2-4.92 (s, 2H), 3.99 (d, J = 17.1 Hz, [(2R,3S,4R,5S,6R)-3,4,5- 2H),3.89-3.78 (m, 4H), trihydroxy-6- 3.75-3.66 (m, 4H), 3.61 (td, J = 9.1,4.4 Hz, (hydroxymethyl)tetrahydropyran- 2H). 2-yl]ethynyl]oxazol-4-yl]ethynyl]tetrahydropyran-3,4,5- triol 130 (2R,3S,4R,5S,6R)-2- (400MHz, CD3OD) δ 9.65 (s, 1H), 502.26 (hydroxymethyl)-6-[2-[8-[2- 8.66 (s,1H), 8.25 (s, 1H), 8.00 (d, J = 7.7 Hz, [(2R,3S,4R,5S,6R)-3,4,5- 1H),7.88 (d, J = 7.3 Hz, trihydroxy-6- 1H), 5.06 (d, J = 7.0 Hz, 2H),(hydroxymethyl)tetrahydropyran- 4.16-4.08 (m, 2H), 3.97 (dd, J = 9.4,3.2 Hz, 2-yl]ethynyl]-5- 2H), 3.94-3.83 (m, 4H),isoquinolyl]ethynyl]tetrahydropyran- 3.75 (dd, J = 11.6, 5.8 Hz, 2H),3.67 (td, 3,4,5-triol J = 9.1, 2.3 Hz, 2H). 131 (2R,3S,4R,5S,6R)-2- (400MHz, CD3OD) δ 7.77 (s, 2H), 509.28 (hydroxymethyl)-6-[2-[4-[2- 4.98 (d,J = 2.1 Hz, 2H), [(2R,3S,4R,5S,6R)-3,4,5- 4.11-4.08 (m, 2H), 4.05 (dd, J= 9.3, 3.3 Hz, trihydroxy-6- 2H), 3.94-3.85 (m, 4H), 3.75 (dd, J = 12.0,(hydroxymethyl)tetrahydropyran- 5.9 Hz, 2H), 3.67 (t, J = 9.5 Hz,2-yl]ethynyl]-2,1,3- 2H). benzothiadiazol-7-yl]ethynyl]tetrahydropyran-3,4,5- triol 132 (2R,3S,4R,5S,6R)-2- (400MHz, CD3OD) δ 7.78 (s, 2H), 513.24 (hydroxymethyl)-6-[2-[6-[2- 4.90 (d,J = 2.1 Hz, 2H), [(2R,3S,4R,5S,6R)-3,4,5- 4.04-4.00 (m, 2H), 3.93 (dd, J= 9.3, 3.3 Hz, trihydroxy-6- 2H), 3.86 (dd, J = 11.5, 2.0 Hz, 2H),(hydroxymethyl)tetrahydropyran- 3.80 (ddd, J = 9.4, 5.7, 2.0 Hz, 2H),2-yl]ethynyl]thieno[3,2- 3.73 (dd, J = 11.5, 5.7 Hz, 2H), b]thiophen-3-3.64 (t, J = 9.4 Hz, 2H). yl]ethynyl]tetrahydropyran-3,4,5- triol 133(2R,3S,4R,5S,6R)-2- (400 MHz, CD3OD) δ 7.46 (d, J = 0.9 Hz, 513.28(hydroxymethyl)-6-[2-[5-[2- 2H), 4.90 (s, 2H), 3.99 (d, J = 2.2 Hz,[(2R,3S,4R,5S,6R)-3,4,5- 2H), 3.90-3.82 (m, 4H), trihydroxy-6- 3.73 (dq,J = 11.8, 6.0 Hz, 4H), (hydroxymethyl)tetrahydropyran- 3.66-3.58 (m,2H). 2-yl]ethynyl]thieno[3,2- b]thiophen-2-yl]ethynyl]tetrahydropyran-3,4,5- triol 134 (2R,3S,4R,5S,6R)-2- (400MHz, CD3OD) δ 8.16 (s, 1H), 482.39 (hydroxymethyl)-6-[2-[4-methoxy- 7.55(s, 1H), 4.89 (s, 1H), 4.01 (d, J = 2.7 Hz,5-[2-[(2R,3S,4R,5S,6R)-3,4,5- 2H), 3.96 (dd, J = 9.8, 2.8 Hz,trihydroxy-6- 1H), 3.92 (s, 3H), (hydroxymethyl)tetrahydropyran-3.91-3.79 (m, 5H), 3.73 (dt, J = 10.1, 7.1 Hz, 2-yl]ethynyl]-2- 3H),3.63 (q, J = 9.1 Hz, 2H). pyridyl]ethynyl]tetrahydropyran- 3,4,5-triol135 (2R,3S,4R,5S,6R)-2- N/A 453.28 (hydroxymethyl)-6-[2-[5-[2-[(2R,3S,4R,5S,6R)-3,4,5- trihydroxy-6- (hydroxymethyl)tetrahydropyran-2-yl]ethynyl]pyrazin-2- yl]ethynyl]tetrahydropyran-3,4,5- triol 136(2R,3S,4R,5S,6R)-2- N/A 501.28 (hydroxymethyl)-6-[2-[4-[2-[(2R,3S,4R,5S,6R)-3,4,5- trihydroxy-6- (hydroxymethyl)tetrahydropyran-2-yl]ethynyl]-1- naphthyl]ethynyl]tetrahydropyran- 3,4,5-triol 137(2R,3S,4R,5S,6R)-2- N/A 501.28 (hydroxymethyl)-6-[2-[8-[2-[(2R,3S,4R,5S,6R)-3,4,5- trihydroxy-6- (hydroxymethyl)tetrahydropyran-2-yl]ethynyl]-1- naphthyl]ethynyl]tetrahydropyran- 3,4,5-triol 138(2R,3S,4R,5S,6R)-2- N/A 457.36 (hydroxymethyl)-6-[2-[5-[2-[(2R,3S,4R,5S,6R)-3,4,5- trihydroxy-6- (hydroxymethyl)tetrahydropyran-2-yl]ethynyl]-2- thienyl]ethynyl]tetrahydropyran- 3,4,5-triol N/A meansnot availablePreparation of Compound 139 (Method A)

(2R,3S,4R,5S,6R)-2-(hydroxymethyl)-6-[3-[4-[3-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]phenyl]phenyl]phenyl]tetrahydropyran-3,4,5-triol

Step I:[(2R,3R,4R,5R,6R)-3,4,5-triacetoxy-6-[3-[4-[3-[(2R,3R,4R,5R,6R)-3,4,5-triacetoxy-6-(acetoxymethyl)tetrahydropyran-2-yl]phenyl]phenyl]phenyl]tetrahydropyran-2-yl]methylacetate

A mixture of Intermediate 0 (101 mg, 0.188 mmol), 1-bromo-4-iodobenzene(86 mg, 0.304 mmol), Siliacat DPP-Pd (72 mg, 0.018 mmol), cesiumcarbonate (135 mg, 0.414 mmol) in CH₃CN (1.9 mL) in a microwave vial ismicrowaved for 10 minutes at 100° C. The resulting mixture is dilutedwith 4 mL of EtOAc, filtered on Celite and evaporated to dryness. Theresidue is used as is for the next step.

Step II: Compound 139

[(2R,3R,4R,5R,6R)-3,4,5-triacetoxy-6-[3-[4-[3-[(2R,3R,4R,5R,6R)-3,4,5-triacetoxy-6-(acetoxymethyl)tetrahydropyran-2-yl]phenyl]phenyl]phenyl]tetrahydropyran-2-yl]methylacetate from Step I is deprotected as previously described usingMeOH/MeONa mixture. ¹H NMR (400 MHz, CD₃OD) δ 7.83 (d, J=1.5 Hz, 2H),7.73 (s, 4H), 7.59 (ddd, J=5.7, 3.8, 2.0 Hz, 2H), 7.46 (dd, J=4.2, 1.6Hz, 4H), 5.04 (d, J=3.7 Hz, 2H), 4.49 (t, J=3.4 Hz, 2H), 3.85 (m, 4H),3.75 (t, J=7.8 Hz, 2H), 3.64 (dd, J=7.9, 3.1 Hz, 2H), 3.55 (ddd, J=7.8,5.6, 4.1 Hz, 2H).

Preparation of Compound 140 (Method A)

Methyl2-[2,5-bis[3-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]phenyl]phenyl]acetate

Compound 140 is prepared according to the procedure described forCompound 139 but using methyl 2-(2,5-dibromophenyl)acetate as startingmaterial. ¹HNMR (400 MHz, CD3OD) δ 7.82 (s, 1H), 7.63-7.56 (m, 3H), 7.50(d, J=7.7 Hz, 1H), 7.48-7.41 (m, 4H), 7.34-7.30 (m, 1H), 7.23 (d, J=7.5Hz, 1H), 5.04 (d, J=3.7 Hz, 1H), 5.01 (d, J=3.6 Hz, 1H), 4.49 (t, J=3.5Hz, 1H), 4.45 (t, J=3.4 Hz, 1H), 3.87-3.81 (m, 4H), 3.78-3.73 (m, 2H),3.70 (s, 2H), 3.66-3.60 (m, 2H), 3.58 (s, 3H), 3.57-3.49 (m, 2H). ESI-MSm/z (M+1)⁺ 627.4

Preparation of Compound 141 (Method F)

(2R,3S,4R,5S,6R)-2-(hydroxymethyl)-6-[2-[4-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]phenyl]ethynyl]tetrahydropyran-3,4,5-triol

Step I:(2R,3R,4R,5R,6R)-2-(4-(((2R,3R,4R,5R,6R)-6-(hydroxymethyl)-3,4,5-tris((triisopropylsilyl)oxy)tetrahydro-2H-pyran-2-yl)ethynyl)phenyl)-6-((pivaloyloxy)methyl)tetrahydro-2H-pyran-3,4,5-triyltris(2,2-dimethylpropanoate)

A vial is charged with Intermediate N (100 mg, 0.153 mmol), IntermediateP (113 mg, 0.168 mmol) in DMF (2.500 mL). The resulting mixture isdegassed. PdCl₂(dppf)₂-CH₂Cl₂ (15.8 mg, 0.0193 mmol) and CuI (8.7 mg,0.046 mmol) are added, degassed again then Et₃N (64 μL, 0.46 mmol) isadded. The vial is capped and stirred at 90° C. overnight. The mixtureis passed through celite and residual solvents are removed under reducedpressure. The crude mixture is used as in the next step without furtherpurification. (201 mg, 0.163 mmol)

Step II:(2R,3R,4R,5R,6R)-2-((pivaloyloxy)methyl)-6-(4-(((2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)ethynyl)phenyl)tetrahydro-2H-pyran-3,4,5-triyltris(2,2-dimethylpropanoate)

To a solution of(2R,3R,4R,5R,6R)-2-(4-(((2R,3R,4R,5R,6R)-6-(hydroxymethyl)-3,4,5-tris((triisopropylsilyl)oxy)tetrahydro-2H-pyran-2-yl)ethynyl)phenyl)-6-((pivaloyloxy)methyl)tetrahydro-2H-pyran-3,4,5-triyltris(2,2-dimethylpropanoate)from Step I (187 mg, 0.152 mmol) in THF (2.1 mL) is added TBAF (607 μLof 1 M, 0.607 mmol). The resulting suspension is stirred overnight atRT. Further diluted with THF (1.5 mL) and treated with Amberlyst Ca²⁺(500 mg) and Amberlyst A-15 H (500 mg) (prewashed, H₂O and THF). Thesuspension is stirred for 1.5h, then filtered and washed with portionsof MeOH. The solvents are removed and the crude mixture is used in thenext step without purification.

Step III: Compound 141

To the crude mixture of(2R,3R,4R,5R,6R)-2-((pivaloyloxy)methyl)-6-(4-(((2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)ethynyl)phenyl)tetrahydro-2H-pyran-3,4,5-triyltris(2,2-dimethylpropanoate) from Step II in MeOH (2 mL) is added MeONa(607 μL of 0.5 M, 0.304 mmol). The resulting mixture is stirred at RTovernight. AcOH (17 μL, 0.30 mmol) is added and the resulting mixture isconcentrated under vacuo. The residue is purified by reverse phase HPLCto afford the title compound (7.6 mg). ¹H NMR (400 MHz, CD₃OD) δ7.51-7.40 (m, 4H), 4.93 (d, J=4.0 Hz, 1H), 4.38-4.29 (m, 1H), 3.98 (dd,J=3.3, 2.1 Hz, 1H), 3.91 (dd, J=9.3, 3.3 Hz, 1H), 3.88-3.76 (m, 4H),3.76-3.68 (m, J=5.6, 3.4 Hz, 2H), 3.62 (t, J=9.5 Hz, 1H), 3.56 (dd,J=7.7, 3.1 Hz, 1H), 3.47 (td, J=6.9, 3.5 Hz, 1H). ESI-MS m/z calc.426.41, found (M+1)⁺ 427.39.

Preparation of Compounds 142-145 and 148

Compounds 142-145, 148 are prepared according to the procedure describedfor Compound 141

LCMS Compound IUPAC name ¹H-NMR m/z (M + H)⁺ 142 (2R,3S,4R,5S,6R)-2-(400 MHz, CD3OD) δ 7.46 (d, J = 8.6 Hz, 441.39 (hydroxymethyl)-6-[2-[3-1H), 7.27 (d, J = 6.7 Hz, 2H), methyl-4-[(2R,3S,4R,5S,6R)- 5.07 (d, J =7.2 Hz, 1H), 4.14 (dd, J = 7.1, 3.2 Hz, 3,4,5-trihydroxy-6- 1H),4.05-3.97 (m, 2H), (hydroxymethyl)tetrahydropyran- 3.97-3.89 (m, 2H),3.88-3.76 (m, 3H), 3.72 (dd, J = 11.6, 2- 5.3 Hz, 2H), 3.65-3.57 (m,2H), yl]phenyl]ethynyl]tetrahydropyran- 2.41 (s, 3H). 3,4,5-triol 143(2R,3S,4R,5S,6R)-2- (400 MHz, CD3OD) δ 7.45-7.35 (m, 441.39(hydroxymethyl)-6-[2-[2- 2H), 7.27 (d, J = 7.9 Hz, 1H),methyl-4-[(2R,3S,4R,5S,6R)- 4.93-4.88 (m, 2H), 4.35 (t, J = 3.5 Hz, 1H),3,4,5-trihydroxy-6- 4.03-3.98 (m, 1H), 3.94 (dd, J = 9.3, 3.3 Hz,(hydroxymethyl)tetrahydropyran- 1H), 3.89-3.77 (m, 4H), 3.77-3.69 (m, 2-2H), 3.63 (t, J = 9.4 Hz, 1H), 3.55 (dd, J = 7.8,yl]phenyl]ethynyl]tetrahydropyran- 3.0 Hz, 1H), 3.46 (dd, J = 10.3, 6.9Hz, 3,4,5-triol 1H), 2.43 (s, 3H). 144 (2R,3S,4R,5S,6R)-2-[2-[2- (400MHz, CD3OD) δ 7.47 (t, J = 7.6 Hz, 445.41 fluoro-4-[(2R,3S,4R,5S,6R)-1H), 7.30 (t, J = 9.8 Hz, 2H), 4.89 (d, 3,4,5-trihydroxy-6- J = 7.5 Hz,2H), 4.23 (t, J = 3.6 Hz, 1H), (hydroxymethyl)tetrahydropyran- 4.04-3.97(m, 1H), 3.95-3.82 (m, 3H), 2-yl]phenyl]ethynyl]-6- 3.82-3.69 (m, 4H),3.63 (t, J = 9.2 Hz, (hydroxymethyl)tetrahydropyran- 1H), 3.58 (dd, J =7.4, 2.7 Hz, 1H), 3,4,5-triol 3.51 (s, 1H). 145 (2R,3S,4R,5S,6R)-2- (400MHz, CD3OD) δ 7.35 (d, J = 7.9 Hz, 457.36 (hydroxymethyl)-6-[2-[2- 1H),7.19 (s, 1H), 6.98 (d, J = 8.0 Hz, methoxy-4- 1H), 4.92 (d, J = 3.9 Hz,1H), 4.35 (t, [(2R,3S,4R,5S,6R)-3,4,5- 1H), 4.05-3.93 (m, J = 7.6, 3.2Hz, 2H), trihydroxy-6- 3.86 (s, 3H), 3.84-3.80 (m, 3H),(hydroxymethyl)tetrahydropyran- 3.76-3.60 (m, 3H), 3.55 (dd, J = 7.8,3.1 Hz, 2- 1H), 3.49 (dd, J = 12.6, 4.9 Hz, 1H),yl]phenyl]ethynyl]tetrahydropyran- 2.06 (s, 1H). 3,4,5-triol 148(2R,3S,4R,5S,6R)-2- (400 MHz, CD3OD) δ 7.50 (m, 1H), 441.35(hydroxymethyl)-6-[2-[2- 7.37 (dd, J = 7.7, 1.3 Hz, 1H), 7.17 (t,methyl-3- J = 7.8 Hz, 1H), 5.11 (d, J = 7.1 Hz, [(2R,3S,4R,5S,6R)-3,4,5-1H), 4.89 (d, J = 2.1 Hz, 1H), trihydroxy-6- 4.15 (dd, J = 7.1, 3.2 Hz,1H), 4.01 (m, (hydroxymethyl)tetrahydropyran- 2H), 3.94 (m, 2H), 3.84(m, 3H), 2- 3.73 (m, 2H), 3.64 (d, J = 9.4 Hz, 1H),yl]phenyl]ethynyl]tetrahydropyran- 3.60 (m, 1H), 2.56 (s, 3H).3,4,5-triolPreparation of Compound 146 (Method D)

(2R,3S,4R,5S,6R)-2-(hydroxymethyl)-6-[2-[2-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]-1H-indol-6-yl]ethynyl]tetrahydropyran-3,4,5-triol

2,5-diiodoaniline (37 mg, 0.11 mmol) and Pd(PPh₃)₄ (12 mg, 0.011 mmol)are charged in a glass vial, the vial is capped, degassed (vacuum thenN₂, 3×) and a solution of Intermediate M (400 μL of a 0.53 M solution inDMF, 0.22 mmol) is added followed by DIPEA (300 μL, 1.72 mmol). Degassedagain then transferred to a preheated oil bath (60° C.) and stirredovernight. The resulting mixture is filtered and purified by reversephase HPLC. Fractions containing the desired material are combined andfreeze-dried to afford the title compound (4.2 mg, 8% yield) as a paleyellow fluffy solid. ¹H NMR (400 MHz, CD₃OD) δ 7.49-7.42 (m, 2H), 7.09(dd, J=8.3, 1.3 Hz, 1H), 6.42 (t, J=0.9 Hz, 1H), 5.25-5.13 (m, 1H), 4.88(d, J=2.0 Hz, 1H), 4.50 (dd, J=3.3, 2.2 Hz, 1H), 4.07-3.96 (m, 2H),3.93-3.84 (m, 3H), 3.83-3.71 (m, 3H), 3.71-3.60 (m, 2H), 3.35-3.23 (m,1H). ESI-MS m/z calc. 465.16348, found (M+1)⁺ 466.47.

Preparation of Compound 150 (Method A)

(2R,2′R,3S,3′S,4R,4′R,5S,5′S,6R,6′R)-6,6′-(2-fluoro-3′-methyl-[1,1′-biphenyl]-4,4′-diyl)bis(2-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol)

To a solution of[(2R,3R,4R,5R,6R)-6-(4-bromo-3-fluoro-phenyl)-3,4,5-tris(2,2-dimethylpropanoyloxy)tetrahydropyran-2-yl]methyl2,2-dimethylpropanoate (90.2 mg, 0.134 mmol; prepared according to theprocedure described for Intermediate N but using1-bromo-2-fluoro-4-iodobenzene as starting material) and Intermediate R(96.0 mg, 0.134 mmol) in 2-methylTHF (2.7 mL) is sequentially added3-(2-dicyclohexylphosphanylphenyl)-2,4-dimethoxy-benzenesulfonic acid(Sodium salt) (13.8 mg, 0.0268 mmol), Pd(OAc)₂ (3.0 mg, 0.013 mmol)(premixed in 0.25 mL of MeTHF), K₂CO₃ (335 μL of 2M, 0.669 mmol) and themixture is stirred at 40° C. overnight. The resulting mixture is dilutedwith AcOEt (10 mL) and water (5 mL), layers are separated and theaqueous layer is back extracted with 10 mL of AcOEt. The combinedorganic layers are dried over Na₂SO₄ and concentrated to dryness. Theresidue is dissolved in MeOH (2 mL) and MeONa (1.0 mL of 0.5 M in MeOH,0.5 mmol) is added and the mixture is stirred overnight at RT. AcOH(30.5 μL, 0.536 mmol) is added, the mixture is concentrated to drynessand the residue is purified by reverse phase preparative HPLC to affordthe title compound (12.4 mg, 18%). ¹H NMR (400 MHz, CD₃OD) δ 7.52 (d,J=8.0 Hz, 1H), 7.46 (t, J=8.1 Hz, 1H), 7.34 (m, 4H), 5.14 (d, J=6.5 Hz,1H), 4.97 (d, J=4.1 Hz, 1H), 4.37 (dd, J=4.1, 3.1 Hz, 1H), 4.26 (dd,J=6.4, 3.3 Hz, 1H), 3.99 (m, 2H), 3.83 (m, 3H), 3.75 (m, 2H), 3.57 (m,3H), 2.49 (s, 3H). ESI-MS m/z calc. 510.51, found (M+1)⁺ 511.42.

Preparation of Compounds 147, 149 and 151

Compounds 147, 149 and 151are prepared according to the proceduredescribed for Compound 150

LCMS Compound IUPAC name ¹H-NMR m/z (M + H)⁺ 147 (2R,3S,4R,5S,6R)-2-(400 MHz, CD3OD) δ 7.44 (d, J = 8.0 Hz, 522.25 (hydroxymethyl)-6-[3-1H), 7.26 (m, 4H), 7.05 (dt, methoxy-4-[3-methyl-4- J = 8.0, 1.2 Hz,1H), 5.12 (d, J = 6.0 Hz, [(2R,3S,4R,5S,6R)-3,4,5- 1H), 5.00 (d, J = 3.4Hz, 1H), trihydroxy-6- 4.46 (t, J = 3.3 Hz, 1H), 4.29 (dd, J = 6.1,(hydroxymethyl)tetrahydropyran- 3.3 Hz, 1H), 4.01 (dd, J = 6.2, 2- 3.3Hz, 1H), 3.96 (dd, J = 11.9, yl]phenyl]phenyl]tetrahydropyran- 6.9 Hz,1H), 3.83 (m, 2H), 3.79 (s, 3,4,5-triol 3H), 3.78 (t, J = 2.0 Hz, 1H),3.74 (m, 1H), 3.70 (d, J = 8.1 Hz, 1H), 3.61 (dd, J = 8.1, 3.1 Hz, 1H),3.52 (ddd, J = 6.8, 5.1, 3.5 Hz, 2H), 2.46 (s, 3H). 149(2R,3S,4R,5S,6R)-2- (400 MHz, CD3OD) δ 7.49 (d, J = 7.2 Hz, N/A(hydroxymethyl)-6-[3-methyl- 1H), 7.42 (m, 1H), 7.37 (m, 4-[3-methyl-4-1H), 7.31 (dd, J = 8.1, 1.8 Hz, 1H), [(2R,3S,4R,5S,6R)-3,4,5- 7.16 (d, J= 7.9 Hz, 1H), 7.11 (d, J = 7.8 Hz, trihydroxy-6- 1H), 5.13 (dd, J =6.3, 5.2 Hz, (hydroxymethyl)tetrahydropyran- 1H), 4.98 (d, J = 3.4 Hz,1H), 2- 4.46 (t, J = 3.3 Hz, 1H), 4.28 (dt, J = 6.2,yl]phenyl]phenyl]tetrahydropyran- 3.6 Hz, 1H), 3.99 (m, 2H), 3,4,5-triol3.83 (m, 3H), 3.76 (m, 2H), 3.61 (dd, J = 8.2, 3.1 Hz, 1H), 3.59-3.46(m, 2H), 2.48 (s, 3H), 2.24 (s, 3H). 151 (2R,3S,4R,5S,6R)-2- (400 MHz,CD3OD) δ 7.65 (m, N/A (hydroxymethyl)-6-[4-[3- 3H), 7.52 (d, J = 8.1 Hz,2H), methoxy-4- 7.24 (dd, J = 8.1, 1.7 Hz, 1H), 7.08 (d, J = 1.7 Hz,[(2R,3S,4R,5S,6R)-3,4,5- 1H), 5.00 (d, J = 3.5 Hz, trihydroxy-6- 1H),4.46 (t, J = 3.3 Hz, 1H), (hydroxymethyl)tetrahydropyran- 3.83 (d, J =4.6 Hz, 2H), 3.74 (t, J = 8.1 Hz, 2- 1H), 3.60 (dd, J = 8.2, 3.1 Hz,yl]phenyl]phenyl]tetrahydropyran- 1H), 3.56 (s, 3H), 3.48 (dt, J = 8.1,3,4,5-triol 4.7 Hz, 1H).Preparation of Compound 152 (Modified Method B)

(2R,3S,4R,5S,6R)-2-(hydroxymethyl)-6-[2-methyl-4-[2-[3-methyl-4-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]phenyl]ethynyl]phenyl]tetrahydropyran-3,4,5-triol

Step I:[(2R,3R,4R,5R,6R)-3,4,5-tris(2,2-dimethylpropanoyloxy)-6-[2-methyl-4-(2-trimethylsilylethynyl)phenyl]tetrahydropyran-2-yl]methyl2,2-dimethylpropanoate

In a glass vial charged with Intermediate Q (259 mg, 0.389 mmol),Pd(dppf)Cl₂—CH₂Cl₂ (33 mg, 0.040 mmol) and CuI (21 mg, 0.11 mmol),capped and flushed with N₂ is added DMF (2.6 mL),ethynyl(trimethyl)silane (275 μL, 1.95 mmol) and Et₃N (270 μL, 1.94mmol). The reaction mixture is transferred to a preheated (80° C.) oilbath and stirred overnight. The reaction mixture is cooled down to RTand diluted with EtOAc and saturated aqueous NH₄Cl (10 mL each). Theorganic layer is separated, washed with saturated aqueous NH₄Cl (2×5mL), brine (5 mL), dried over Na₂SO₄ and passed through a 2 g silicacartridge, using EtOAc. The filtrate is concentrated then purifiedBiotage™ SNAP 10 g silica cartridge, using a gradient of EtOAc in Hex,0-20% as eluent. The desired fractions are combined and concentrated,providing the title compound (246 mg, 93% yield) as a yellow foam.

Step II:(2R,3S,4R,5S,6R)-2-(4-ethynyl-2-methyl-phenyl)-6-(hydroxymethyl)tetrahydropyran-3,4,5-triol

To a solution of[(2R,3R,4R,5R,6R)-3,4,5-tris(2,2-dimethylpropanoyloxy)-6-[2-methyl-4-(2-trimethylsilylethynyl)phenyl]tetrahydropyran-2-yl]methyl2,2-dimethylpropanoate from Step I (244 mg, 0.355 mmol) in MeOH (2.5 mL)is added MeONa in MeOH (2.1 mL of 0.5 M, 1.1 mmol) and the mixture istransferred to a preheated (60° C.) oil bath. After stirring for 3h, thereaction mixture is cooled down to RT and treated with prewashed Dowex50WX4-400 resin, filtered and washed with portions of MeOH. The combinedfiltrates are concentrated and purified by flash chromatography on abond elute 5 g silica cartridge, using a gradient of MeOH in CH₂Cl₂, 0to 20% as eluent. Combined fractions are concentrated affording thetitle compound (90 mg, 91% yield) as an amber foamy solid.

Step III:[(2R,3R,4R,5R,6R)-3,4,5-tris(2,2-dimethylpropanoyloxy)-6-[2-methyl-4-[2-[3-methyl-4-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]phenyl]ethynyl]phenyl]tetrahydropyran-2-yl]methyl2,2-dimethylpropanoate

In a glass vial charged with(2R,3S,4R,5S,6R)-2-(4-ethynyl-2-methyl-phenyl)-6-(hydroxymethyl)tetrahydropyran-3,4,5-triol(35.0 mg, 0.12 mmol), Intermediate Q (70.0 mg, 0.105 mmol), CuI (10 mg,0.053 mmol) and Pd(dppf)Cl₂— CH₂Cl₂ (10 mg, 0.014 mmol), capped andflushed with N₂ is added DMF (0.7 mL) and DIEA (55 μL, 0.32 mmol). Thevial is degassed and transferred to a preheated oil bath (90° C.), andstirred for 1h and slowly cooled to RT and left overnight. The reactionmixture is diluted with EtOAc and saturated NH₄Cl (10 mL each) andfiltered. The layers are separated, the aqueous layer is back extractedwith EtOAc (5 mL). The combined organic extracts are washed withsaturated NH₄Cl solution (5 mL), brine (5 mL), dried over Na₂SO₄,filtered and concentrated. The crude product is purified on Biotage™SNAP 10 g silica cartridge, using a gradient of MeOH in CH₂Cl₂, 0-20% aseluent. The combined fractions are concentrated to provide the titlecompound (28 mg, 31% yield) as a pinkish brown solid.

Step IV: Compound 152

Deprotection of[(2R,3R,4R,5R,6R)-3,4,5-tris(2,2-dimethylpropanoyloxy)-6-[2-methyl-4-[2-[3-methyl-4-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]phenyl]ethynyl]phenyl]tetrahydropyran-2-yl]methyl2,2-dimethylpropanoate from Step III (28 mg, 0.32 mmol) using theprotocol described in step II provides the title compound (17 mg, 94%yield) as an off-white fluffy solid (after redissolving the crudeproduct in H₂O/MeCN mixture (20% MeCN) and freeze-drying). ¹H NMR (400MHz, CD₃OD) δ 7.49 (d, J=8.6 Hz, 2H), 7.34 (d, J=7.2 Hz, 4H), 5.11 (d,J=7.0 Hz, 2H), 4.20 (dd, J=7.0, 3.3 Hz, 2H), 4.04 (dd, J=11.9, 7.4 Hz,2H), 3.99 (dd, J=5.5, 3.3 Hz, 2H), 3.85 (dd, J=5.5, 4.3 Hz, 2H), 3.75(dd, J=11.9, 3.8 Hz, 2H), 3.66-3.57 (m, 2H), 2.46 (s, 6H). ESI-MS m/zcalc. 530.2152. found 531.55 (M+1)⁺.

Preparation of Compound 155 (Method A)

(2R,3S,4R,5S,6R)-2-(hydroxymethyl)-6-[2-methyl-4-[2-methyl-6-[3-methyl-4-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]phenyl]-3-pyridyl]phenyl]tetrahydropyran-3,4,5-triol

Step I:(2R,3R,4R,5R,6R)-2-(4-(5-bromo-6-methylpyridin-2-yl)-2-methylphenyl)-6-((pivaloyloxy)methyl)tetrahydro-2H-pyran-3,4,5-triyltris(2,2-dimethylpropanoate)

A 5 ml microwave vial is charged with Intermediate R (150 mg, 0.209mmol), 3,6-dibromo-2-methyl-pyridine (43.8 mg, 0.174 mmol), SiliacatDPP-Pd (70 mg, 0.017 mmol) and Cs₂CO₃ (171 mg, 0.523 mmol) in CH₃CN (2.0mL). The vial is capped under nitrogen and irradiated at 130° C. for 30min in the microwave. The reaction mixture is diluted with EtOAc passedon a 500 mg silica cartridge and eluted with EtOAc. The resultingmixture was concentrated under reduced pressure the residue (159 mg) isused as such in the next step.

[(2R,3R,4R,5R,6R)-3,4,5-tris(2,2-dimethylpropanoyloxy)-6-[2-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]tetrahydropyran-2-yl]methyl2,2-dimethylpropanoate is prepared according to the procedure describedfor Intermediate B and N

Step II

A 5 ml microwave vial is charged with a crude mixture of[(2R,3R,4R,5R,6R)-6-[4-(5-bromo-6-methyl-2-pyridyl)-2-methyl-phenyl]-3,4,5-tris(2,2-dimethylpropanoyloxy)tetrahydropyran-2-yl]methyl2,2-dimethylpropanoate (130 mg, 0.1709 mmol), Intermediate R (123 mg,0.171 mmol), Siliacat DPP-Pd (68.4 mg, 0.0171 mmol) and Cs₂CO₃ (167.0mg, 0.513 mmol) in CH₃CN (2.7 mL). The vial is capped under nitrogen andirradiated at 130° C. for 30 min in the microwave. The reaction mixtureis diluted with EtOAc passed on a 500 mg silica cartridge and elutedwith EtOAc. The residual mixture was concentrated under reduced pressureto afford the crude mixture of the title compound, used as such in thenext step

Step III: Compound 155

The residue is dissolved in MeOH (1.30 mL). MeONa (46.2 mg, 0.855 mmol)is added to the solution and the latter is allowed to stir for 48 h atRT. AcOH (58 μL, 1.03 mmol) is added to the mixture and the solvents areremoved under reduced pressure. The residue is then purified byreverse-phase HPLC to afford the title compound (12.6 mg, 12%). ¹H NMR(400 MHz, CD3OD) δ 7.83-7.72 (m, J=8.2 Hz, 2H), 7.66 (dd, J=18.0, 8.1Hz, 2H), 7.58 (dd, J=7.9, 5.3 Hz, 2H), 7.26-7.18 (m, 2H), 5.16 (d, J=6.7Hz, 2H), 4.27 (dd, J=6.6, 2.8 Hz, 2H), 4.07-3.92 (m, 4H), 3.89-3.81 (m,2H), 3.76 (dt, J=11.9, 3.5 Hz, 2H), 3.65-3.53 (m, J=11.1, 8.0, 4.0 Hz,2H), 2.54 (s, 3H), 2.52 (s, 3H), 2.51 (s, 3H). LC-MS: m/z=598.59 (M+H⁺).

Preparation of Compounds 153, 154 and 157

Compounds 153, 154 and 157 prepared according to the procedure describedfor Compound 155

Com- LCMS m/z pound IUPAC name ¹H-NMR (M + H)⁺ 153 (2R, 3S, 4R, 5S,6R)-2- (400 MHz, CD3OD) δ 9.09 585.57 (hydroxymethyl)-6-[2- (s, 2H),7.92 (m, 4H), 7.63 methyl-4-[5[3-methyl-4- (d, J = 8.7 Hz, 2H), 5.15[(2R, 3S, 4R, 5S, 6R)- (d, J = 6.8 Hz, 2H), 4.24 3,4,5-trihydroxy-6-(dd, J = 6.8, 3.2 Hz, 2H), (hydroxymethyl)tetra- 4.02 (m, 4H), 3.85 (dd,hydropyran-2- J = 5.7, 4.3 Hz, 2H), 3.75 yl]phenyl]pyrazin- (dd, J =11.9, 3.7 Hz, 2H), 2-yl]phenyl]tetrahydro- 3.61 (dt, J = 7.7, 4.0 Hz,pyran-3,4,5-triol 2H), 2.55 (s, 6H). 154 (2R, 3S, 4R, 5S, 6R)-2- (400MHz, CD3OD) δ 584.59 (hydroxymethyl)-6-[2- 7.95 (d, J = 7.2 Hz, 4H),methyl-4-[6-[3-methyl-4- 7.88 (dd, J = 8.5, 7.1 Hz, [(2R, 3S, 4R, 5S,6R)- 1H), 7.77 (d, J = 7.6 Hz, 3,4,5-trihydroxy-6- 2H), 7.59 (d, J = 8.7Hz, (hydroxymethyl)tetra- 2H), 5.16 (d, J = 6.5 Hz,hydropyran-2-yl]phenyl]- 2H), 4.28 (dd, J = 6.4, 2-pyridyl]phenyl]tetra-3.3 Hz, 2H), 4.08-3.92 (m, hydropyran-3,4,5-triol 4H), 3.84 (t, 2H),3.76 (dd, J = 11.9, 3.7 Hz, 2H), 3.58 (dt, J = 8.3, 4.1 Hz, 2H), 2.56(s, 6H). 157 (2R, 3S, 4R, 5S, 6R)-2- 584.63 (hydroxymethyl)-6-[2-methyl-4-[6-[3-methyl-4- [(2R, 3S, 4R, 5S, 6R)-3, 4,5-trihydroxy-6-(hydroxymethyl)tetra- hydropyran-2-yl]phenyl]- 3-pyridyl]phenyl]tetra-hydropyran-3,4,5-triolPreparation of Compound 156

2R,3S,4R,5S,6R)-2-(hydroxymethyl)-6-[4-[2-[3-methyl-4-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]phenyl]cyclopropyl]phenyl]tetrahydropyran-3,4,5-triol

Step I:(2R,3R,4R,5R,6R)-2-(4-((E)-2-(6-methyl-4,8-dioxo-1,3,6,2-dioxazaborocan-2-yl)vinyl)phenyl)-6-((pivaloyloxy)methyl)tetrahydro-2H-pyran-3,4,5-triyltris(2,2-dimethylpropanoate)

To a vial containing[(2R,3R,4R,5R,6R)-3,4,5-triacetoxy-6-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]tetrahydropyran-2-yl]methylacetate (200 mg, 0.374 mmol),2-[(E)-2-bromovinyl]-6-methyl-1,3,6,2-dioxazaborocane-4,8-dione (118 mg,0.449 mmol) is added under a nitrogen atmosphere C₃₄H₂₈Cl₂FeP₂Pd (27.4mg, 0.0374 mmol) and K₃PO₄ (238.4 mg, 1.123 mmol). CH₃CN (5.5 mL) isadded and the vial is sealed and allowed to stir at RT for 3 days. Themixture is passed on a silica gel pad, the solvents are removed toafford a crude mixture of the title compound (104 mg, 0.1765 mmol), usedas such in the next step.

Step II:(2R,3R,4R,5R,6R)-2-(4-((E)-3-methyl-4-((2R,3R,4R,5R,6R)-3,4,5-tris(pivaloyloxy)-6-((pivaloyloxy)methyl)tetrahydro-2H-pyran-2-yl)styryl)phenyl)-6-((pivaloyloxy)methyl)tetrahydro-2H-pyran-3,4,5-triyltris(2,2-dimethylpropanoate)

To a vial containing[(2R,3R,4R,5R,6R)-6-(4-bromo-2-methyl-phenyl)-3,4,5-tris(2,2-dimethylpropanoyloxy)tetrahydropyran-2-yl]methyl2,2-dimethylpropanoate (119.0 mg, 0.1777 mmol),[(2R,3R,4R,5R,6R)-3,4,5-triacetoxy-6-[4-[(E)-2-(6-methyl-4,8-dioxo-1,3,6,2-dioxazaborocan-2-yl)vinyl]phenyl]tetrahydropyran-2-yl]methylacetate (104.7 mg, 0.1777 mmol), C₃₄H₂₈Cl₂FeP₂Pd (130.0 mg, 0.1777mmol), K₃PO₄ (113.2 mg, 0.5331 mmol) is added under a nitrogenatmosphere in CH₃CN (1.2 mL) and water (251 μL). The vial is sealed andallowed to stir at RT for 24 h. The mixture is passed on a silica pad,(CH₂Cl₂ and EtOAc) the solvents are removed and the residue is purifiedusing (0-80%) EtOAc in Hex as solvents to afford the title compound(46.5 mg, 26%).

Step III:[(2R,3R,4R,5R,6R)-3,4,5-tris(2,2-dimethylpropanoyloxy)-6-[4-[(E)-2-[3-methyl-4-[(2R,3R,4R,5R,6R)-3,4,5-tris(2,2-dimethylpropanoyloxy)-6-(2,2-dimethylpropanoyloxymethyl)tetrahydropyran-2-yl]phenyl]vinyl]phenyl]tetrahydropyran-2-yl]methyl2,2-dimethylpropan

To a solution of[(2R,3R,4R,5R,6R)-3,4,5-tris(2,2-dimethylpropanoyloxy)-6-[2-methyl-4-[(E)-2-[4-[(2R,3R,4R,5R,6R)-3,4,5-triacetoxy-6-(acetoxymethyl)tetrahydropyran-2-yl]phenyl]vinyl]phenyl]tetrahydropyran-2-yl]methyl2,2-dimethylpropanoate (46.0 mg, 0.0450 mmol) and palladium acetate (3.0mg, 0.014 mmol) in CH₂Cl₂ (1.2 mL) at 0° C. is added a solution ofdiazomethane (9.0 mL of 0.5 M, 4.5 mmol) dropwise and the solution isstirred until complete conversion. LCMS shows complete conversion toproduct. The resulting mixture is filtered over celite and the filtrateis concentrated under reduced pressure to afford as a crude mixture ofthe title compound used as such in the next step.

Step IV: Compound 156

The residue is diluted in MeOH (1.4 mL) and MeONa (540 μL of 0.5 M,0.270 mmol) is added. The solution is allowed to stir for 72 h at RT.AcOH (13 μL, 0.23 mmol) is added, the volatiles are removed underreduced pressure and the residue is purified by reverse-phase HPLC toafford the title compound (4.2 mg, 17%). ¹H NMR (400 MHz, CD3OD) δ 7.34(dd, J=18.6, 8.3 Hz, 3H), 7.14 (d, J=8.3 Hz, 2H), 7.01-6.86 (m, J=5.1Hz, 2H), 5.06 (d, J=6.0 Hz, 1H), 4.94 (d, J=3.2 Hz, 1H), 4.42 (t, J=3.2Hz, 1H), 4.24 (dd, J=5.8, 3.2 Hz, 1H), 4.01-3.86 (m, J=18.7, 9.0, 5.0Hz, 2H), 3.79 (t, J=4.6 Hz, 3H), 3.77-3.67 (m, J=16.5, 5.9 Hz, 2H), 3.56(dd, J=8.3, 3.1 Hz, 1H), 3.50-3.37 (m, 2H), 2.41 (s, 3H), 2.17-2.01 (m,J=14.5, 6.6 Hz, 2H), 1.52-1.29 (m, 2H). LC-MS: m/z=533.52 (M+H⁺).

Preparation of Compound 158 (Method D)

(2R,3S,4R,5S,6R)-2-[2-[3-benzyl-4-[2-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]ethynyl]phenyl]ethynyl]-6-(hydroxymethyl)tetrahydropyran-3,4,5-triol

Compound 158 is prepared according to the procedure described for 59. ¹HNMR (400 MHz, CD3OD) δ 7.46 (d, J=7.9 Hz, 1H), 7.36-7.24 (m, 4H),7.22-7.14 (m, 3H), 4.94-4.87 (m, 1H), 4.85 (d, J=1.9 Hz, 1H), 4.14 (s,2H), 4.00-3.97 (m, 1H), 3.97-3.94 (m, 1H), 3.92-3.68 (m, 8H), 3.65 (d,J=8.5 Hz, 1H), 3.60 (d, J=9.3 Hz, 1H). LC-MS: m/z (M+H)⁺=541.5

Preparation of Compound 159 (Method B)

(2R,3S,4R,5S,6R)-2-(hydroxymethyl)-6-[2-methyl-4-[(E)-2-[3-methyl-4-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]phenyl]vinyl]phenyl]tetrahydropyran-3,4,5-triol

Compound 159 is prepared according to the procedure described for 156.¹H NMR (400 MHz, CD3OD) δ 7.43 (d, J=8.0 Hz, 2H), 7.37 (d, J=10.8 Hz,4H), 7.11 (s, 2H), 5.09 (d, J=6.2 Hz, 2H), 4.24 (dd, J=6.2, 3.2 Hz, 2H),4.04-3.92 (m, J=11.9, 7.5 Hz, 4H), 3.82 (t, J=5.4 Hz, 2H), 3.74 (dd,J=11.9, 3.6 Hz, 2H), 3.58-3.50 (m, 2H), 2.46 (s, 6H).

Preparation of Compound 160 (Method B)

(2R,3S,4R,5S,6R)-2-(hydroxymethyl)-6-[2-methyl-4-[2-[3-methyl-4-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]phenyl]cyclopropyl]phenyl]tetrahydropyran-3,4,5-triol

Compound 160 is prepared according to the procedure described for 156.(400 MHz, CD3OD) δ 7.37-7.27 (m, J=8.6 Hz, 2H), 6.99-6.89 (m, J=4.7 Hz,4H), 5.06 (d, J=5.9 Hz, 2H), 4.24 (dd, J=5.8, 3.3 Hz, 2H), 4.01-3.87 (m,J=18.7, 9.0, 5.1 Hz, 4H), 3.79 (t, J=5.8 Hz, 2H), 3.73 (dd, J=11.8, 3.6Hz, 2H), 3.49-3.41 (m, 2H), 2.41 (s, 6H), 2.12-2.02 (m, J=7.3 Hz, 2H),1.44-1.34 (m, 2H).

Preparation of Compound 162 (Method D)

(2R,2′R,3S,3′S,4R,4′R,5S,5′S,6R,6′R)-6,6′-((2′,3′,5′,6′-tetrahydrospiro[fluorene-9,4′-pyran]-2,7-diyl)bis(ethyne-2,1-diyl))bis(2-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol)

Step I: Compound 162 Crude

A mixture of 2,7-dibromospiro[fluorene-9,4′-tetrahydropyran] (12.00 g,30.45 mmol) and Intermediate M (16.02 g, 85.14 mmol) in DMF (168.0 mL)is degased for 5 minutes by bubbling nitrogen in the reaction mixture.Pd(dppf)Cl₂— CH₂Cl₂ (1.680 g, 2.296 mmol) and CuI (1.685 g, 8.849 mmolare added and nitrogen is bubbled one more time in the reaction mixturesfor 5 minutes. Diisopropyl ethyl amine (42.0 mL, 241 mmol) is then addedand the final mixture is stirred under nitrogen atmosphere at 100° C.for 1 h. The resulting reaction mixture is cooled to 35° C. and water(336.0 mL) is added dropwise. The resulting deep red mixture is stirredovernight at RT and the resulting precipitate is filtered (ML 1), washedwith 100 ml of water. The resulting crude orange solid is transferred ina 250 ml RBF and triturated in 60 ml of ethanol for 30 minutes. Theresulting solid is isolated by filtration and dried in a vacuum ovenovernight at 45° C. To further purify the title compound, the latter isper-acetylated (Step 2), submitted to flash chromatography andde-acetylated (Step 3).

Step II:[(2R,3R,4R,5R,6R)-3,4,5-triacetoxy-6-[2-[7-[2-[(2R,3R,4R,5R,6R)-3,4,5-triacetoxy-6-(acetoxymethyl)tetrahydropyran-2-yl]ethynyl]spiro[fluorene-9,4′-tetrahydropyran]-2-yl]ethynyl]tetrahydropyran-2-yl]methylAcetate

To the crude solid from Step 1 (13.77 g) is added DMAP (276 mg, 2.26mmol) and pyridine (69 mL). The resulting mixture (brown suspension) iscooled to 0° C. in an ice/water bath and Ac₂O (25.6 mL, 271 mmol) isadded dropwise over 10 minutes keeping temperature below 20° C. Thereaction mixture is then stirred at RT for one h. The resulting darkbrown solution is diluted with CH₂Cl₂ (100 mL) and water (75 mL),stirred for 15 min, then 2N HCl (˜475 mL) is added and stirred for 5 minwith ice/water bath to control exotherm. The aqueous solution isseparated, back-extracted with CH₂Cl₂ (2×75 mL) and the combined organicextracts are washed once again with 2N HCl (75 mL, added brine to helpseparation), dried over Na₂SO₄, filtered and concentrated to afford20.78 g of an orange glassy solid. The solid is dissolved in a minimumamount of CH₂Cl₂, adsorbed on 75 ml of silica gel and purified usingBiotage™ SNAP 340 g silica cartridge, using a gradient of Hex/EtOAC(100% Hex 1 CV, 0-85% EA/hex 16 CV, 85% EA/hex 4 CV). The fractionscontaining the desired compound are combined and treated with 4.5 g ofSiliaMet® Thiol overnight at RT to remove traces of palladium. Theresulting mixture is the filtered and concentrated to dryness to yield16.87 g of the title compound.

Step III: Compound 162

[(2R,3R,4R,5R,6R)-3,4,5-triacetoxy-6-[2-[7-[2-[(2R,3R,4R,5R,6R)-3,4,5-triacetoxy-6-(acetoxymethyl)tetrahydropyran-2-yl]ethynyl]spiro[fluorene-9,4′-tetrahydropyran]-2-yl]ethynyl]tetrahydropyran-2-yl]methylacetate. From Step II (15.37 g, 16.27 mmol) is dissolved in MeOH (344mL) and MeONa (290 μL of 25% w/w, 1.30 mmol in MeOH) is added. Final pHreached is 9. The reaction is stirred at RT for 18 h. The resultingsuspension (white solid) is filtered and the solid washed with 8 volumesof MeOH. The white solid is dried in a vacuum oven at 40° C. overnightto afford the title compound as a beige solid (9.38 g, 15.03 mmol)containing 1.6% of a mono-acetate impurity. ¹H NMR (400 MHz, DMSO) δ7.94 (d, J=7.9 Hz, 2H), 7.88 (d, J=1.4 Hz, 2H), 7.50 (dd, J=7.8, 1.3 Hz,2H), 4.98 (d, J=4.3 Hz, 2H), 4.81 (d, J=5.9 Hz, 2H), 4.78-4.67 (m, 4H),4.50 (t, J=6.0 Hz, 2H), 4.09 (q, J=5.2 Hz, 0H), 4.00 (dd, J=7.1, 3.8 Hz,4H), 3.89-3.80 (m, 2H), 3.80-3.65 (m, 4H), 3.60 (ddd, J=8.8, 6.3, 2.0Hz, 2H), 3.48 (dt, J=12.0, 6.2 Hz, 2H), 3.40 (td, J=9.3, 5.9 Hz, 2H),3.16 (d, J=5.3 Hz, 0H), 2.03 (s, OH), 1.81 (dd, J=7.3, 3.8 Hz, 4H).LC-MS: m/z=609.57 (M+H)⁺

Preparation of Compounds 161, 162 and 164

Compounds 161, 162 and 164 are prepared according to the proceduredescribed for 120 using commercially available dibromo fluorenederivatives

Com- LCMS m/z pound IUPAC name ¹H-NMR (M + H)⁺ 161 (2R, 3S, 4R, 5S, 6R)-N/A 589.16 2-[2-[5,5-dioxo-7-[2- [(2R, 3S, 4R, 5S, 6R)-3,4,5-trihydroxy-6- (hydroxymethyl)tetra- hydropyran-2-yl]ethynyl]dibenzo- thiophen-3-yl]ethynyl]- 6-(hydroxymethyl)tetra-hydropyran-3,4,5-triol 163 2,7-bis[2-[(2R, 3S, 4R, (400 MHz, CD3OD) δ568.43 5S, 6R)-3,4,5-tri- 8.48 (d, J = 33.2 Hz, 1H), hydroxy-6-(hydroxy-7.84-7.66 (m, 3H), 7.52 methyl)tetrahydro- (dd, J = 27.9, 7.7 Hz, 2H),pyran-2-yl]ethynyl] 4.87 (s, 2H), 4.05-4.00 (m, fluoren-9-one oxime 2H),3.98-3.92 (m, 2H), 3.91-3.80 (m, 4H), 3.73 (dd, J = 11.5, 5.7 Hz, 2H),3.63 (t, J = 9.4 Hz, 2H). 164 (2R, 3S, 4R, 5S, 6R)- (400 MHz, CD3OD) δ616.43 2-(hydroxymethyl)-6- 8.31 (s, 2H), 7.71-7.64 [2-[9-phenyl-6-[2-(m, 2H), 7.59-7.48 (m, [(2R, 3S, 4R, 5S, 6R)- 5H), 7.31 (d, J = 8.6 Hz,3,4,5-trihydroxy-6- 2H), 4.87 (s, 2H), 4.07-3.97 (hydroxymethyl)tetra-(m, 4H), 3.88 (d, J = 9.9 hydropyran-2- Hz, 4H), 3.75 (dd, J = 11.9,yl]ethynyl]carbazol-3- 6.2 Hz, 2H), 3.70-3.61 yl]ethynyl]tetrahydro- (m,2H). pyran-3,4,5-triolPreparation of Compounds 165 to 170Compounds 165 to 170 are prepared according to the procedure describedfor 113 using the appropriate Intermediates.

Compound/ LCMS m/z Intermediate IUPAC name ¹H-NMR (M + H)⁺ 165 (2R, 3S,4R, 5S, 6R)-2-[2- (400 MHz, CD3OD) δ 7.67 (d, 597.32[9-hydroxy-9-isopropyl-7- J = 7.6 Hz, 2H), 7.55 (s, 2H), [2-[(2R, 3S,4R, 5S, 6R)- 7.47 (d, J = 7.7 Hz, 2H), 4.59 3,4,5-trihydroxy-6- (s, 2H),4.01 (s, 2H), 3.95 (d, (hydroxymethyl)tetrahydrop- J = 9.1 Hz, 2H),3.90-3.79 (m, yran-2-yl]ethynyl]fluoren- 4H), 3.73 (dd, J = 11.7, 5.6Hz, 2-yl]ethynyl]-6- 2H), 3.63 (t, J = 9.5 Hz, 2H),(hydroxymethyl)tetrahydrop- 2.41 (dt, J = 13.7, 6.8 Hz, 1H),yran-3,4,5-triol 0.76 (d, J = 6.7 Hz, 6H). 166 (2R, 3S, 4R, 5S,6R)-2-[2- (400 MHz, CD3OD) δ 7.68 (d, 583.31 [9-ethyl-9-hydroxy-7-[2- J= 7.7 Hz, 2H), 7.55 (s, 2H), [(2R, 3S, 4R, 5S, 6R)-3,4,5- 7.47 (d, J =7.8 Hz, 2H), 4.59 trihydroxy-6- (s, 2H), 4.05-3.99 (m, 2H),(hydroxymethyl)tetrahydrop- 3.95 (dd, J = 9.3, 3.2 Hz, 2H),yran-2-yl]ethynyl]fluoren-2- 3.91-3.79 (m, 4H), 3.73 (dd, yl]ethynyl]-6-J = 11.6, 5.7 Hz, 2H), 3.63 (t, J = (hydroxymethyl)tetrahydrop- 9.4 Hz,2H), 2.15 (dd, J = 15.3, yran-3,4,5-triol 7.9 Hz, 2H), 0.42 (t, J = 7.3Hz, 3H). 167 (2R, 3S, 4R, 5S, 6R)-2- (400 MHz, CD3OD) δ 7.75 (d, 631.19(hydroxymethyl)-6-[2-[9- J = 7.9 Hz, 2H), 7.47 (d, J =hydroxy-9-phenyl-7-[2- 7.9 Hz, 2H), 7.32-7.18 (m, [(2R, 3S, 4R, 5S,6R)-3,4,5- 7H), 4.82 (d, J = 2.0 Hz, 2H), trihydroxy-6- 3.98-3.94 (m,2H), 3.91-3.80 (hydroxymethyl)tetrahydrop- (m, 4H), 3.79-3.73 (m, 2H),yran-2-yl]ethynyl]fluoren-2- 3.69 (dd, J = 11.5, 6.0 Hz, 2H),yl]ethynyl]tetrahydropyran- 3.58 (t, J = 9.4 Hz, 2H). 3,4,5-triol 168(2R, 3S, 4R, 5S, 6R)-2- (400 MHz, CD3OD) δ 7.67 (d, 597.32(hydroxymethyl)-6-[2-[9- J = 7.8 Hz, 2H), 7.55 (s, 2H),hydroxy-9-propyl-7-[2- 7.47 (d, J = 8.0 Hz, 2H), 4.59 [(2R, 3S, 4R, 5S,6R)-3,4,5- (s, 2H), 4.02 (d, J = 2.6 Hz, trihydroxy-6- 2H), 3.95 (d, J =9.0 Hz, 2H), (hydroxymethyl)tetrahydrop- 3.90-3.80 (m, 4H), 3.73 (dd,yran-2-yl]ethynyl]fluoren-2- J = 11.6, 5.7 Hz, 2H), 3.63 (t, J =yl]ethynyl]tetrahydropyran- 9.4 Hz, 2H), 2.13-2.05 (m, 3,4,5-triol 2H),0.74 (s, 5H). 169 (2R, 3S, 4R, 5S, 6R)-2-[2- (400 MHz, CD3OD) δ 7.68 (d,611.4 [9-hydroxy-9-isobutyl-7-[2- J = 7.9 Hz, 2H), 7.56 (s, 2H), [(2R,3S, 4R, 5S, 6R)-3,4,5- 7.48 (dd, J = 7.9, 1.4 Hz, 2H), trihydroxy-6-4.88 (s, 2H), 4.02 (t, J = 2.6 (hydroxymethyl)tetrahydrop- Hz, 2H), 3.95(dt, J = 9.3, 3.3 yran-2-yl]ethynyl]fluoren-2- Hz, 2H), 3.90-3.80 (m,4H), yl]ethynyl]-6- 3.73 (dd, J = 11.5, 5.6 Hz, 2H),(hydroxymethyl)tetrahydrop- 3.63 (t, J = 9.5 Hz, 2H), 2.12yran-3,4,5-triol (d, J = 6.0 Hz, 2H), 0.95 (td, J = 13.0, 6.6 Hz, 1H),0.51 (d, J = 6.7 Hz, 6H). 170 (2R, 3S, 4R, 5S, 6R)-2-[2- (400 MHz,CD3OD) δ 7.74 (d, 661.19 [9-hydroxy-9-(3- J = 7.9 Hz, 2H), 7.47 (d, J =methoxyphenyl)-7[2- 7.9 Hz, 2H), 7.30 (s, 2H), 7.13 [(2R, 3S, 4R, 5S,6R)-3,4,5- (t, J = 8.1 Hz, 1H), 6.96 (d, J = trihydroxy-6- 1.5 Hz, 1H),6.78 (dd, J = 8.0, (hydroxymethyl)tetrahydrop- 2.2 Hz, 1H), 6.67 (d, J =7.8 yran-2-yl]ethynyl]fluoren-2- Hz, 1H), 4.83 (s, 2H), 3.99-3.94yl]ethynyl]-6- (m, 2H), 3.92-3.80 (m, (hydroxymethyl)tetrahydrop- 4H),3.79-3.65 (m, 7H), 3.58 yran-3,4,5-triol (t, J = 9.5 Hz, 2H).Preparation of Compound 171 (Method D)

(2R,3S,4R,5S,6R)-2-(hydroxymethyl)-6-[2-[7-[2-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]ethynyl]-1H-indazol-4-yl]ethynyl]tetrahydropyran-3,4,5-triol

To a reaction tube charged with commercially available4,7-dibromo-1H-indazole (35.0 mg, 0.127 mmol) and Pd(PPh₃)₄ (18.0 mg,0.0156 mmol), capped and degassed (vacuum then nitrogen flush, twice) isadded Intermediate M as a solution in DMF (500 μL of 0.53 M, 0.265 mmoland DIPEA (500 μL). The reaction tube is degassed again, transferred toa preheated (80° C.) oil bath and stirred for 48h. After cooling down toRT, the reaction mixture is passed through a 200 mg Si-DMT cartridge,rinsed with portions of MeOH and purified by reverse phase HPLC. Thefraction is freeze-dried, providing the title compound (12.6 mg, 20%yield) as a fluffy white solid. ¹H NMR (400 MHz, CD3OD) δ 8.21 (s, 1H),7.50 (d, J=7.5 Hz, 1H), 7.28 (d, J=7.5 Hz, 1H), 5.02 (d, J=2.1 Hz, 1H),5.00 (d, J=2.0 Hz, 1H), 4.16-4.08 (m, 2H), 4.04-3.83 (m, 6H), 3.82-3.72(m, 2H), 3.71-3.59 (m, 2H). ESI-MS m/z: 491.44 (M+1)⁺

Preparation of Compound 172:

1,4-bis[3-methyl-4-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]phenyl]piperazine-2,5-dione

Step I:[(2R,3R,4R,5R,6R)-3,4,5-tris(2,2-dimethylpropanoyloxy)-6-[2-methyl-4-[4-[3-methyl-4-[(2R,3R,4R,5R,6R)-3,4,5-tris(2,2-dimethylpropanoyloxy)-6-(2,2-dimethylpropanoyloxymethyl)tetrahydropyran-2-yl]phenyl]-2,5-dioxo-piperazin-1-yl]phenyl]tetrahydropyran-2-yl]

To a reaction tube loaded with Intermediate Q (213 mg, 0.318 mmol),piperazine-2,5-dione (12.6 mg, 0.110 mmol), CuI (7.8 mg, 0.0410 mmol)and K₂CO₃ (50.0 mg, 0.362 mmol), capped and degassed (vacuum thennitrogen flush, 3×) is added degassed DMF (1 mL) followed byN,N′-dimethylethane-1,2-diamine (8.0 μL, 0.075 mmol). The reaction tubeis degassed again and transferred to a preheated (110° C.) oil bath andstirred overnight. The reaction mixture is cooled down to RT, dilutedwith EtOAc (5 mL), and washed sequentially with saturated aqueous NH₄Clsolution (2×5 mL), H₂O (2 mL), brine (2 mL), dried over Na₂SO₄ andpassed through a 500 mg silica cartridge using EtOAc. The filtrate isconcentrated then purified by flash chromatography on a Biotage™ SNAP 10g silica cartridge, using a gradient of EtOAc in Hex, 0-100% as eluent.The fractions are combined and concentrated to provide the titlecompound (40 mg, 28% yield) as a white crystalline solid.

Step II: Compound 172

To a solution of the intermediate from Step I above (38.0 mg, 0.0294mmol) in MeOH (1.4 mL) is added a solution of MeONa in MeOH (90 μL of0.5 M, 0.0450 mmol). The reaction mixture is stirred at 60° C. for 4 h,then passed through a prewashed (MeOH) 1 g SCX-2 cartridge, washing withportions of MeOH (3×1 mL). The combined filtrates are concentrated,redissolved in a H₂O/MeCN mixture (20% MeCN) and freeze-dried, affordingthe title compound (19 mg, 96% yield) as a fluffy white solid. ¹H NMR(400 MHz, CD3OD) δ 7.60 (d, J=9.1 Hz, 2H), 7.30-7.21 (m, 4H), 5.12 (d,J=7.0 Hz, 2H), 4.52 (s, 4H), 4.20 (dd, J=7.0, 3.2 Hz, 2H), 4.05 (dd,J=11.9, 7.4 Hz, 2H), 4.00 (dd, J=5.4, 3.3 Hz, 2H), 3.88-3.81 (m, 2H),3.75 (dd, J=11.9, 3.8 Hz, 2H), 3.63 (dt, J=7.6, 3.9 Hz, 2H), 2.50 (s,6H). ESI-MS m/z: 619.6 (M+1)⁺

Preparation of Compounds 173 and 174

Compounds 173 and 174 are prepared according to the procedure describedfor 59 using commercially available starting material. Reaction mixturesare stirred 1h at 100° C.

LCMS m/z Compound IUPAC name ¹H-NMR (M + H)⁺ 173 (2R, 3S, 4R, 5S,6R)-2-[2-[9-[3- (400 MHz, CD3OD) δ 8.24 641.36(dimethylamino)-2-hydroxy- (s, 2H), 7.63-7.53 (m, 4H),propyl]-6-[2-[(2R, 3S, 4R, 5S, 4.89 (s, 2H), 4.44-4.34 (m,6R)-3,4,5-trihydroxy-6- 3H), 4.01 (ddd, J = 12.5,(hydroxymethyl)tetrahydropyran- 6.2, 2.7 Hz, 4H), 3.882-yl]ethynyl]carbazol-3- (ddd, J = 9.4, 4.3, 2.1 Hz, yl]ethynyl]-6- 4H),3.75 (dd, J = 12.2, (hydroxymethyl)tetrahydropyran- 6.3 Hz, 2H), 3.65(t, J = 3,4,5-triol 9.3 Hz, 2H), 2.72 (s, 6H). 174 (2R, 3S, 4R, 5S,6R)-2-[2-[9- (400 MHz,CD3OD) δ 8.22 614.32 (2,3-dihydroxypropyl)-6-[2-(d, J = 1.2 Hz, 2H), 7.55 [(2R, 3S, 4R, 5S, 6R)-3,4,5- (dt, J = 8.5, 4.9Hz, 4H), trihydroxy-6- 4.88 (s, 2H), 4.42 (ddd, J =(hydroxymethyl)tetrahydropyran- 22.6, 14.6, 5.8 Hz, 1H),2-yl]ethynyl]carbazol-3- 4.05-3.99 (m, 5H), 3.89 yl]ethynyl]-6- (ddd, J= 9.1, 5.5, 2.2 Hz, (hydroxymethyl)tetrahydropyran- 4H), 3.75 (dd, J =12.3, 6.3 3,4,5-triol Hz, 2H), 3.65 (t, J = 9.2 Hz, 2H), 3.57 (dd, J =5.5, 2.1 Hz, 2H), 3.46 (dt, J = 3.2, 1.5 Hz, 1H).Preparation of Compound 175 (Method D)

Ethyl2-[2,7-bis[2-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]ethynyl]carbazol-9-yl]acetate

Step I: Compound 175

To a mixture of Intermediates AG1 (200 mg, 0.49 mmol) and M (1.96 mL of0.53 M, 1.036 mmol), Pd(dppf)Cl₂— CH₂Cl₂ (26.2 mg, 0.032 mmol), CuI(27.8 mg, 0.146 mmol) in DMF (800.0 μL) is degassed (vacuum/N₂). To theresulting mixture is added DIPEA (678 μL, 3.89 mmol), degassed. Thereaction mixture is heated at 100° C. for 2 h 45 min under N₂,concentrated under high vacuum, dissolved in DMSO (1.5 mL), loaded ontoC18 Samplet, The residue is purified on 50 g C-18 silica gel cartridgeon Isolera™ purification system with a gradient of CH₃CN in water(10%-45%, 12.5 CV) as eluent to afford after concentrating the titlecompound (140 mg, 42%) as light yellow solid. ¹H NMR (400 MHz, CD₃OD) δ8.06 (d, J=8.1 Hz, 2H), 7.57 (s, 2H), 7.32 (dd, J=8.1, 1.2 Hz, 2H), 5.20(s, 2H), 4.89 (d, J=2.1 Hz, 2H), 4.20 (q, J=7.1 Hz, 2H), 4.03 (dd,J=3.2, 2.2 Hz, 2H), 3.98 (dd, J=9.3, 3.3 Hz, 2H), 3.91-3.84 (m, 4H),3.78-3.71 (m, 2H), 3.64 (t, J=9.5 Hz, 2H), 1.24 (t, J=7.1 Hz, 3H).ESI-MS m/z calc. 625.21594. found 626.56 (M+1)⁺.

Preparation of Compound 176 (Method D)

2-[2,7-Bis[2-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]ethynyl]carbazol-9-yl]aceticAcid

Step I: Compound 176

To mixture of ethyl2-[2,7-bis[2-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]ethynyl]carbazol-9-yl]acetate(Compound 175, 140 mg, 0.205 mmol) in EtOH (2 mL) and H₂O (1.5 mL) isadded aqueous NaOH (250 μL of 10% w/v, 0.6250 mmol). The final mixtureis heated in a sealed tube at 80° C. for 1 h (LC-MS showed cleanproduct). Reaction mixture is quenched with DOWEX 50WX4 hydrogen formresin until pH 4-5, it formed the gel, diluted with water-methanol (10mL) until the gel is dissolved, filtered off, concentrated, dissolved inwater and CH₃CN, lyophilized to afford the title compound (112 mg,88.7%) as light yellow solid. ¹H NMR (400 MHz, CD₃OD) δ 8.04 (d, J=8.1Hz, 2H), 7.56 (s, 2H), 7.29 (d, J=8.1 Hz, 2H), 4.06-3.96 (m, 4H),3.92-3.82 (m, 4H), 3.80-3.70 (m, 2H), 3.64 (t, J=9.3 Hz, 2H), 2.64 (s,4H). (Cl—H protons are underneath of water peak). ESI-MS m/z calc.597.18463. found 598.59 (M+1)⁺.

Preparation of Compound 177 (Method D)

2-[2,7-bis[2-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]ethynyl]carbazol-9-yl]-N-[2-(4-methylpiperazin-1-yl)ethyl]acetamide

To a stirred solution of 2-(4-methylpiperazin-1-yl)ethanamine (7.7 mg,0.054 mmol) and Et₃N (20 μL) in DMF (0.3 mL) is added a premixedsolution of Compound 176 and HATU (23 mg, 0.061 mmol) in DMF (0.4 mL).Reaction mixture is stirred at RT for 3 h and the mixture is purifieddirectly by reverse phase HPLC to afford the title compound (16 mg, 40%yield). ¹H NMR (400 MHz, CD₃OD) δ 8.38 (brs, 1H), 8.11 (d, J=8.1 Hz,2H), 7.63 (s, 2H), 7.36 (dd, J=8.1, 1.2 Hz, 2H), 5.04 (s, 2H), 4.90 (d,J=2.1 Hz, 2H), 4.03 (dd, J=3.2, 2.2 Hz, 2H), 3.96 (dd, J=9.3, 3.3 Hz,2H), 3.91-3.81 (m, 4H), 3.73 (dd, J=11.3, 5.8 Hz, 2H), 3.63 (t, J=9.4Hz, 2H), 2.64 (s, 3H), 2.46 (t, J=6.0 Hz, 2H). ¹H NMR (400 MHz, DMSO-D₆)δ 8.17 (d, J=8.1 Hz, 2H), 8.08 (t, J=5.6 Hz, 1H), 7.64 (s, 2H),7.30-7.25 (m, 2H), 5.06 (s, 2H), 4.76 (d, J=2.0 Hz, 2H), 3.83 (s, 2H),3.77-3.65 (m, 4H), 3.63-3.55 (m, 2H), 3.46 (dd, J=11.7, 6.2 Hz, 2H),3.39 (t, J=9.3 Hz, 2H), 3.16 (dd, J=12.1, 6.2 Hz, 2H), 2.40-2.16 (m,10H), 2.11 (s, 3H). ESI-MS m/z calc. 722.3163. found 723.71 (M+1)⁺.

Preparation of Compounds 178 to 181.

Compounds 178 to 181 are prepared according to the procedure describedfor 177 using appropriate commercially available starting material.

LCMS m/z Compound IUPAC name ¹H-NMR (M + H)⁺ 178 2-[2,7-bis[2-[(2R, 3S,4R, 5S, (400 MHz, CD₃OD) δ 8.10 710.69 6R)-3,4,5-trihydroxy-6- (d, J =8.1 Hz, 2H), 7.60 (s, (hydroxymethyl)tetrahydropyran- 2H), 7.35 (d, J =8.1 Hz, 2H), 2-yl]ethynyl]carbazol-9-yl]-N-(2- 5.06 (s, 2H), 4.89 (d, J= 2.1 morpholinoethyl)acetamide Hz, 2H), 4.05-4.00 (m, 2H), 3.96 (dd, J= 9.3, 3.2 Hz, 2H), 3.91-3.82 (m, 4H), 3.74 (dd, J = 11.8, 6.0 Hz, 2H),3.69-3.59 (m, 6H), 3.50-3.40 (m, 2H), 2.81 (s, 6H). [1] 1792-[2,7-bis[2-[(2R, 3S, 4R, 5S, (400 MHz, CD₃OD) δ 8.05 (d, 680.676R)-3,4,5-trihydroxy-6- J = 8.1 Hz, 2H), 7.55 (s, 2H),(hydroxymethyl)tetrahydropyran- 7.30 (d, J = 8.1 Hz, 2H), 5.342-yl]ethynyl]carbazol-9-yl]-1-(4- (s, 2H), 4.03 (dd, J = 3.2, 2.2methylpiperazin-1-yl)ethanone Hz, 2H), 3.97 (dd, J = 9.3, 3.3 Hz, 2H),3.91-3.61 (m, 12H), 2.86-2.78 (m, 2H), 2.70 (s, 2H), 2.52 (s, 3H) andtwo protons under the solvent peak 180 2-[2,7-bis[2-[(2R, 3S, 4R, 5S,(400 MHz, CD₃OD) δ 8.05 (d, 667.36 6R)-3,4,5-trihydroxy-6- J = 8.1 Hz,2H), 7.57 (s, 2H), (hydroxymethyl)tetrahydropyran- 7.30 (d, J = 8.1 Hz,2H), 5.35 2-yl]ethynyl]carbazol-9-yl]-1- (s, 2H), 4.89 (d, J = 2.2 Hz,morpholino-ethanone 2H), 4.05-4.01 (m, 2H), 3.98 (dd, J = 9.3, 3.3 Hz,2H), 3.87 (ddd, J = 11.6, 7.1, 2.2 Hz, 4H), 3.83-3.79 (m, 2H), 3.77-3.69(m, 6H), 3.65 (t, J = 9.5 Hz, 2H), 3.59-3.55 (m, 2H). [1] 1812-[2,7-bis[2-[(2R, 3S, 4R, 5S, (400 MHz, CD₃OD) δ 8.05 625.376R)-3,4,5-trihydroxy-6- (d, J = 8.1 Hz, 2H), 7.55 (s,(hydroxymethyl)tetrahydropyran- 2H), 7.30 (d, J = 8.1 Hz, 2H),2-yl]ethynyl]carbazol-9-yl]-N,N- 5.32 (s, 2H), 4.89 (d, J = 2.1dimethyl-acetamide Hz, 2H), 4.05-4.01 (m, 2H), 3.98 (dd, J = 9.3, 3.2Hz, 2H), 3.91-3.82 (m, 4H), 3.74 (dd, J = 11.6, 5.6 Hz, 2H), 3.64 (t, J= 9.6 Hz, 2H), 3.27 (s, 3H), 2.98 (s, 3H).Preparation of Compound 182 (Method D)

(2R,3S,4R,5S,6R)-2-[2-[9-(2-Hydroxyethyl)-7-[2-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]ethynyl]carbazol-2-yl]ethynyl]-6-(hydroxymethyl)tetrahydropyran-3,4,5-triol

A mixture of Intermediates AG2 (60 mg, 0.15 mmol) and M (588 μL of 0.53M, 0.311 mmol) Pd(dppf)Cl₂— CH₂Cl₂ (7.9 mg, 0.01 mmol), CuI (8.4 mg,0.044 mmol) in DMF (215 μL) is degassed (vacuum/N₂). To the resultingmixture is added DIPEA (204 μL, 1.17 mmol), and degassed. The reactionmixture is heated at 100° C. for 2 h under N₂, passed throughmetal-scavenger cartridge (Si-DMT; Silicycle; SPE-R79030B-06P), washedwith DMF (0.5 mL). The resulting filtrate is concentrated under highvacuum, dissolved in DMSO, purified by reverse phase HPLC to afford thetitle compound (24 mg, 28%) as light yellow solid. ¹H NMR (400 MHz,CD₃OD) δ 8.05 (d, J=8.1 Hz, 2H), 7.68 (s, 2H), 7.29 (d, J=8.1 Hz, 2H),4.91-4.89 (m, 2H), 4.45 (t, J=5.4 Hz, 2H), 4.06-4.02 (m, 2H), 3.99 (dd,J=9.3, 3.3 Hz, 2H), 3.94-3.84 (m, 6H), 3.79-3.71 (m, 2H), 3.65 (t, J=9.4Hz, 2H). ESI-MS m/z calc. 583.2054. found 584.54 (M+1)⁺.

Preparation of Compound 183 (Method D)

(2R,3S,4R,5S,6R)-2-(Hydroxymethyl)-6-[2-[9-[2-(trideuteriomethoxy)ethyl]-7-[2-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]ethynyl]carbazol-2-yl]ethynyl]tetrahydropyran-3,4,5-triol

Compound 183 is prepared according to the procedure described forCompound 182 but using Intermediates AG3 (63 mg, 0.15 mmol) and M (590.0μL of 0.53 M, 0.313 mmol). The title compound is isolated as white solidby filtration after purification by reverse phase HPLC as itprecipitated out of solution (16.7 mg, 19%). ¹H NMR (400 MHz, DMSO-D₆) δ8.15 (d, J=8.1 Hz, 2H), 7.72 (s, 2H), 7.26 (d, J=8.1 Hz, 2H), 4.96 (d,J=4.3 Hz, 2H), 4.82 (d, J=5.7 Hz, 2H), 4.76 (s, 2H), 4.72 (d, J=5.9 Hz,2H), 4.58 (s, 2H), 4.50 (t, J=5.9 Hz, 2H), 3.84 (s, 2H), 3.79-3.55 (m,8H), 3.52-3.34 (m, 4H). ESI-MS m/z calc. 600.23987, found (M+1)⁺601.58.

Preparation of Compound 184 (Method D)

(2R,3S,4R,5S,6R)-2-(hydroxymethyl)-6-[2-[9-tetrahydropyran-4-yl-7-[2-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]ethynyl]carbazol-2-yl]ethynyl]tetrahydropyran-3,4,5-triol

A solution of Intermediates M (889 μL of 0.53 M in DMF, 0.47 mmol) andAG34 (90 mg, 0.22 mmol), Pd(dppf)Cl₂— CH₂Cl₂ (13 mg, 0.016 mmol), CuI(13 mg, 0.068 mmol) is degased (vacuum/N₂). To the resulting mixture isadded DIPEA (320 μL, 1.84 mmol). The final mixture is stirred in asealed tube under N₂ atmosphere at 100° C. for 2h. The reaction iscooled down, filtered over SiliCycle SiliaPrep DMT 200 mg 3 mL SPEcartridge and concentrated in vacuo. The residue is purified by reversephase HPLC to afford the title compound (15 mg, 11%) as a solid. ¹H NMR(400 MHz, CD₃OD) δ 8.06 (d, J=8.1 Hz, 2H), 7.79 (s, 2H), 7.30 (dd,J=8.0, 1.1 Hz, 2H), 4.91 (d, J=2.1 Hz, 2H), 4.22-4.10 (m, 2H), 4.05 (dd,J=3.2, 2.1 Hz, 2H), 3.99 (dd, J=9.3, 3.3 Hz, 2H), 3.93-3.82 (m, 4H),3.81-3.60 (m, 7H), 2.68 (qd, J=12.6, 4.7 Hz, 2H), 1.83 (dd, J=11.8, 3.6Hz, 2H). ESI-MS m/z calc. 623.65. found 624.56 (M+1)⁺.

Preparation of Compounds 185 to 190.

Compounds 185 to 190 are prepared according to the procedure describedfor 184 using Intermediates AG35 to AG40

LCMS m/z Compound IUPAC name ¹H-NMR (M + H)⁺ 185 tert-butyl4-[2,7-bis[2- (400 MHz, CD₃OD) δ 8.06 (d, J = 722.14 [(2R, 3S, 4R, 5S,6R)-3,4,5- 8.0 Hz, 2H), 7.72 (s, 2H), 7.30 trihydroxy-6- (dd, J = 8.1,1.2 Hz, 2H), 4.87 (s, (hydroxymethyl)tetrahydropyran- 2H), 4.78 (s, 1H),4.60 (s, 1H), 2-yl]ethynyl]carbazol-9- 4.32 (s, 2H), 4.03 (dd, J = 3.3,2.2 yl]piperidine-1-carboxylate Hz, 2H), 3.98 (dd, J = 9.3, 3.3 Hz, 2H),3.92-3.82 (m, 4H), 3.79-3.71 (m, 2H), 3.65 (t, J = 9.5 Hz, 2H),3.24-3.17 (m, 2H), 1.54 (s, 9H). Three proton under solvent peaks 186(2R, 3S, 4R, 5S, 6R)-2- (400 MHz, CD₃OD) δ 8.09 (d, J = 625.58(hydroxymethyl)-6-[2-[9-[(3- 8.1 Hz, 2H), 7.67 (s, 2H), 7.33 (d,methyloxetan-3-yl)methyl]-7-[2- J = 8.1 Hz, 2H), 4.90 (d, J = 2.1 [(2R,3S, 4R, 5S, 6R)-3,4,5- Hz, 2H), 4.79-4.68 (m, 2H), 4.49 trihydroxy-6-(s, 2H), 4.30 (d, J = 6.0 Hz, 2H), (hydroxymethyl)tetrahydropyran- 4.04(t, J = 2.7 Hz, 2H), 3.98 (dd, 2-yl]ethynyl]carbazol-2- J = 9.3, 3.4 Hz,2H), 3.92-3.81 yl]ethynyl]tetrahydropyran-3,4,5- (m, 4H), 3.74 (dd, J =11.7, 5.8 triol Hz, 2H), 3.65 (t, J = 9.6 Hz, 2H), 1.43 (s, 3H). 187(2R, 3S, 4R, 5S, 6R)-2- N/A 638.2 (hydroxymethyl)-6-[2-[9-(1-methyl-4-piperidyl)-7-[2- [(2R, 3S, 4R, 5S, 6R)-3,4,5- trihydroxy-6-(hydroxymethyl)tetrahydropyran- 2-yl]ethynyl]carbazol-2-yl]ethynyl]tetrahydropyran-3,4,5- triol 188 (2R, 3S, 4R, 5S, 6R)-2- (400MHz, CD₃OD) δ 8.07 (d, J = 652.6 (hydroxymethyl)-6-[2-[9-[(1- 8.1 Hz,2H), 7.68 (d, J = 1.2 Hz, methyl-4-piperidyl)methyl]-7[2- 2H), 7.31 (dd,J = 8.1, 1.2 Hz, [(2R, 3S, 4R, 5S, 6R)-3,4,5- 2H), 4.90 (d, J = 2.1 Hz,2H), trihydroxy-6- 4.45 (t, J = 7.1 Hz, 2H), 4.03 (dd,(hydroxymethyl)tetrahydropyran- J = 3.3, 2.1 Hz, 2H), 3.97 (dd, J =2-yl]ethynyl]carbazol-2- 9.3, 3.3 Hz, 2H), 3.87 (ddt, J =yl]ethynyl]tetrahydropyran-3,4,5- 10.0, 7.7, 2.3 Hz, 4H), 3.78-3.71triol (m, 2H), 3.65 (t, J = 9.6 Hz, 2H), 3.31 (s, 1H), 3.25 (s, 2H),2.90 (s, 1H), 2.79 (s, 3H), 2.41 (s, 1H), 2.20 (s, 1H), 2.03 (dd, J =11.6, 6.8 Hz, 2H), 1.72 (m, 1H). 189 (2R, 3S, 4R, 5S, 6R)-2- (400 MHz,CD₃OD) δ 8.05 (dd, 654.56 (hydroxymethyl)-6-[2-[9-(2- J = 8.0, 0.7 Hz,2H), 7.68 (dd, J = morpholinoethyl)-7-[2- 1.3, 0.7 Hz, 2H), 7.30 (dd, J= [(2R, 3S, 4R, 5S, 6R)-3,4,5- 8.1, 1.2 Hz, 2H), 4.90 (d, J = 2.1trihydroxy-6- Hz, 2H), 4.51 (t, J = 6.7 Hz, 2H),(hydroxymethyl)tetrahydropyran- 4.04 (dd, J = 3.3, 2.1 Hz, 2H),2-yl]ethynyl]carbazol-2- 3.99 (dd, J = 9.3, 3.3 Hz, 2H),yl]ethynyl]tetrahydropyran-3,4,5- 3.93-3.82 (m, 4H), 3.81-3.70 triol (m,2H), 3.70-3.57 (m, 6H), 2.78 (t, J = 6.7 Hz, 2H), 2.56 (t, J = 4.5 Hz,4H). 190 1-[2-[2,7-bis[2- (400 MHz, CD₃OD) δ 8.07 (d, J = 651.6 [(2R,3S, 4R, 5S, 6R)-3,4,5- 8.1 Hz, 2H), 7.67 (s, 2H), 7.31 (d, trihydroxy-6-J = 8.1 Hz, 2H), 4.90 (d, J = 2.1 (hydroxymethyl)tetrahydropyran- Hz,2H), 4.59 (t, J = 5.7 Hz, 2H), 2-yl]ethynyl]carbazol-9- 4.12-3.95 (m,4H), 3.88 (ddd, yl]ethyl]pyrrolidin-2-one J = 11.8, 7.1, 2.3 Hz, 4H),3.75 (dd, J = 11.7, 5.8 Hz, 2H), 3.70-3.54 (m, 4H), 2.86 (t, J = 7.1 Hz,2H), 2.12 (t, J = 8.1 Hz, 2H), 1.60 (q, J = 7.6 Hz, 2H).Preparation of Compound 191 (Method D)

(2R,3S,4R,5S,6R)-2-(hydroxymethyl)-6-[2-[7′-[2-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]ethynyl]spiro[1,3-dithiane-2,9′-fluorene]-2′-yl]ethynyl]tetrahydropyran-3,4,5-triol

Step I: 2′,7′-dibromospiro[1,3-dithiane-2,9′-fluorene]

To a solution of 2,7-dibromofluoren-9-one (1000 mg, 2.959 mmol) inCH₂Cl₂ (9.863 mL) at RT under argon is added propane-1,3-dithiol (446μL, 4.44 mmol) followed by BF₃.OEt₂ (548 μL, 4.44 mmol). The resultingmixture is stirred at RT for 1 h and then warmed to 60° C. overnight.The reaction mixture is quenched by pouring into saturated aqueousNa₂CO₃, and extracted with EtOAc. The combined organic extracts aredried over Na₂SO₄, filtered and concentrated. During the concentrationprocess a precipitate is formed and it is isolate by filtration toafford the title compound (0.937 g, 2.188 mmol, 73.95%). LC-MS:m/z=428.13 (M+H⁺).

Step II: Compound 191

To a degased mixture of intermediate M (925 μL of 0.53 M, 0.490 mmol),2′,7′-dibromospiro[1,3-dithiane-2,9′-fluorene] from Step I (100 mg,0.2335 mmol), Pd(dppf)Cl₂—CH₂Cl₂ (19.07 mg, 0.02335 mmol), CuI (13.34mg, 0.07005 mmol) in DMF (778.3 μL) is added DIPEA (122 μL, 0.701 mmol).The mixture is stirred in a sealed tube under nitrogen atmosphere at 90°C. overnight. The reaction mixture is filtered over celite cartridge andconcentrated in vacuo. Purification by reverse phase HPLC afford thetitle compound (37.2 mg, 25%). ¹H NMR (400 MHz, CD₃OD) δ 7.95 (s, 2H),7.78 (d, J=7.9 Hz, 2H), 7.53 (dd, J=7.9 Hz, 2H), 4.90 (d, J=1.9 Hz, 3H),4.04 (t, 2H), 3.96 (dd, J=9.4, 3.2 Hz, 2H), 3.90 (d, J=2.0 Hz, 1H),3.89-3.80 (m, 3H), 3.74 (dd, J=11.4, 5.6 Hz, 2H), 3.63 (t, J=9.4 Hz,2H), 2.38-2.28 (m, 2H). LC-MS: m/z=643.51 (M+H⁺).

Preparation of Compound 192 (Method D)

(2R,3S,4R,5S,6R)-2-(Hydroxymethyl)-6-[2-[7-[2-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]ethynyl]spiro[fluorene-9,4′-piperidine]-2-yl]ethynyl]tetrahydropyran-3,4,5-triol

Compound 192 is prepared according to the procedure described forCompound 182 but using Intermediates AG4 (45 mg, 0.08940 mmol) and M(360 μL of 0.53 M, 0.1908 mmol). Half of the resulting crude mixture isdissolved in DMSO (1 mL), purified by reverse phase HPLC. Two drops ofammonia is added to the combined fractions (pH 6-7), followed bylyophilisation (twice) to afford the title compound (11.8 mg, 43%) aslight yellow solid. ¹H NMR (400 MHz, CD₃OD) δ 7.85 (d, J=7.9 Hz, 2H),7.82 (s, 2H), 7.55 (d, J=7.9 Hz, 2H), 4.89 (d, J=2.1 Hz, 2H), 4.05-3.98(m, 2H), 3.93 (dd, J=9.3, 3.3 Hz, 2H), 3.89-3.79 (m, 4H), 3.74 (dd,J=11.4, 5.8 Hz, 2H), 3.67-3.57 (m, 6H), 2.11-2.03 (m, 4H). ESI-MS m/zcalc. 607.24176. found 608.58 (M+1)⁺.

Preparation of Compound 193 (Method D)

(2R,3S,4R,5S,6R)-2-[2-[9,9-Bis(2-hydroxyethyl)-7-[2-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]ethynyl]fluoren-2-yl]ethynyl]-6-(hydroxymethyl)tetrahydropyran-3,4,5-triol

Compound 193 is prepared according to the procedure described forCompound 182 but using Intermediates AG6 (40 mg, 0.097 mmol) and M (390μL of 0.53 M, 0.207 mmol). The title compound is obtained (24 mg, 40%)as white solid after lyophilisation. ¹H NMR (400 MHz, CD₃OD) δ 7.75 (d,J=7.9 Hz, 2H), 7.60 (d, J=0.7 Hz, 2H), 7.47 (dd, J=7.9, 1.4 Hz, 2H),4.88 (d, J=2.3 Hz, 2H), 4.02 (dd, J=3.2, 2.2 Hz, 2H), 3.95 (dd, J=9.3,3.3 Hz, 2H), 3.91-3.80 (m, 4H), 3.74 (dd, J=11.4, 5.5 Hz, 2H), 3.63 (t,J=9.4 Hz, 2H), 2.82-2.70 (m, 4H), 2.39-2.29 (m, 4H). ESI-MS m/z calc.626.2363. found 627.54 (M+1)⁺.

Preparation of Compound 194 (Method D)

(2R,3S,4R,5S,6R)-2-[2-[(3R,4S)-3,4-Dihydroxy-7′-[2-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]ethynyl]spiro[cyclopentane-1,9′-fluorene]-2′-yl]ethynyl]-6-(hydroxymethyl)tetrahydropyran-3,4,5-triol

Compound 194 is prepared according to the procedure described forCompound 182 but using Intermediates AG7 (41 mg, 0.01 mmol) and M (400μL of 0.53 M, 0.213 mmol). Purification by reverse phase HPLC followedby lyophilisation afforded the title compound (31 mg, 50%) as a whitsolid. ¹H NMR (400 MHz, CD₃OD) δ 7.95 (s, 1H), 7.72-7.64 (m, 2H), 7.52(s, 1H), 7.46-7.38 (m, 2H), 4.53-4.41 (m, 2H), 4.06-3.98 (m, 2H),3.99-3.92 (m, 2H), 3.91-3.78 (m, 4H), 3.74 (dd, J=11.3, 5.3 Hz, 2H),3.63 (t, J=9.4 Hz, 2H), 2.32-2.11 (m, 4H). [Two protons for C-1sugaroverlapped with solvent peak). ESI-MS m/z calc. 624.2207. found 625.53(M+1)⁺.

Preparation of Compounds 195 to 199.

Compounds 195-199 are prepared according to the procedure described forCompound 182 but using the appropriate commercially available startingmaterial (for Compound 195) and Intermediates AG27 to AG29 (for Compound196 to 199 respectively)

LCMS m/z Compound IUPAC name ¹H-NMR (M + H)⁺ 195 (2R, 3S, 4R, 5S, 6R)-2-(400 MHz, CD₃OD) δ 7.74-7.66 555.3 (hydroxymethyl)-6-[2-[9-hydroxy- (m,2H), 7.49 (d, J = 7.8 7-[2-[(2R, 3S, 4R, 5S, 6R)-3,4,5- Hz, 1H),7.46-7.42 (m, 1H), trihydroxy-6- 7.36-7.31 (m, 2H), 4.84 (d, J =(hydroxymethyl)tetrahydropyran- 2.3 Hz, 2H), 4.79 (s, 1H),2-yl]ethynyl]-9H-fluoren-2- 4.03-3.97 (m, 2H), 3.93 (td, J =yl]ethynyl]tetrahydropyran-3,4,5- 9.5, 3.1 Hz, 2H), 3.89-3.77 (m, triol4H), 3.77-3.69 (m, 2H), 3.62 (td, J = 9.5, 5.0 Hz, 2H). 196 (2R, 3S, 4R,5S, 6R)-2-[2-[9-(2,3- (400 MHz, CD₃OD) δ 7.65 (dd, 629.3dihydroxypropyl)-9-hydroxy-7- J = 29.2, 5.5 Hz, 4H), 7.47 (d, [2-[(2R,3S, 4R, 5S, 6R)-3,4,5- J = 7.4 Hz, 2H), 4.79 (s, 2H), trihydroxy-6- 4.59(s, 1H), 4.00 (s, 2H), 3.94 (hydroxymethyl)tetrahydro- (d, J = 10.8 Hz,3H), 3.84 (dd, pyran-2-yl]ethynyl]fluoren-2- J = 22.1, 9.9 Hz, 5H), 3.73(dd, yl]ethynyl]-6-(hydroxy- J = 11.1, 5.0 Hz, 2H),methyl)tetrahydropyran-3,4,5- 3.67-3.58 (m, 2H), 2.31-2.21 triol (m,1H), 2.09 (dd, J = 14.1, 3.3 Hz, 1H). 197 (2R, 3S, 4R, 5S,6R)-2-[2-[(4′R, (400 MHz, CD₃OD) δ 7.92 (s, 641.35′S)-4′,5′-dihydroxy-7-[2- 1H), 7.82 (s, 1H), 7.73 (d, J = [(2R, 3S, 4R,5S, 6R)-3,4,5- 7.8 Hz, 1H), 7.67 (d, J = 7.8 Hz, trihydroxy-6- 1H), 7.50(dd, J = 16.5, 7.8 Hz, (hydroxymethyl)tetrahydropyran- 2H), 4.79 (d, J =0.9 Hz, 2H), 2-yl]ethynyl]spiro[fluorene-9,2′- 4.49-4.42 (m, 1H),4.25-4.18 tetrahydropyran]-2-yl]ethynyl]-6 (m, 1H), 4.02 (dd, J = 3.0,2.1 (hydroxymethyl)tetrahydropyran- Hz, 2H), 3.97-3.92 (m, 2H),3,4,5-triol 3.90-3.80 (m, 4H), 3.73 (ddd, (2R, 3S, 4R, 5S,6R)-2-[2-[(4′S, J = 11.6, 5.5, 1.5 Hz, 2H), 3.635′R)-4′,5′-dihydroxy-7-[2- (t, J = 9.4 Hz, 2H), 2.47 (dd, J = [(2R, 3S,4R, 5S, 6R)-3,4,5- 13.2, 10.9 Hz, 1H), 1.74 (dd, trihydroxy-6- J = 13.5,5.1 Hz, 1H). (hydroxymethyl)tetrahydropyran-2-yl]ethynyl]spiro[fluorene-9,2′- tetrahydropyran]-2-yl]ethynyl]-6-(hydroxymethyl)tetrahydropyran- 3,4,5-triol 198 (2R, 3S, 4R, 5S, 6R)-2-(400 MHz, CD₃OD) δ 7.80-7.71 637.33 (hydroxymethyl)-6-[2-[9-(4- (m, 4H),7.52 (d, J = 7.9 methylpiperazin-1-yl)-7-[2- Hz, 2H), 5.00 (s, 1H), 4.84(s, [(2R, 3S, 4R, 5S, 6R)-3,4,5- 2H), 4.02-3.98 (m, 2H), 3.92trihydroxy-6- (ddd, J = 9.4, 3.0, 1.7 Hz, 2H),(hydroxymethyl)tetrahydropyran- 3.87 (d, J = 11.8 Hz, 2H),2-yl]ethynyl]-9H-fluoren-2- 3.84-3.78 (m, 2H), 3.74 (dd, J =yl]ethynyl]tetrahydropyran-3,4,5- 11.4, 5.7 Hz, 2H), 3.64 (t, J = triol9.4 Hz, 2H), 2.92 (s, 3H), 2.76 (s, 4H), 2.58 (s, 4H). 199 (2R, 3S, 4R,5S, 6R)-2- (400 MHz, CD₃OD) δ 7.82 (s, 609.32(hydroxymethyl)-6-[2-[7-[2- 2H), 7.71 (dd, J = 7.8, 3.1 Hz, [(2R, 3S,4R, 5S, 6R)-3,4,5- 2H), 7.50 (d, J = 7.8 Hz, 2H), trihydroxy-6- 4.78 (s,2H), 4.10-4.05 (m, (hydroxymethyl)tetrahydropyran- 2H), 4.05-3.99 (m,2H), 3.95 2-yl]ethynyl]spiro[fluorene-9,2′- (dd, J = 9.3, 3.3 Hz, 2H),tetrahydropyran]-2- 3.91-3.80 (m, 4H), 3.73 (ddd,yl]ethynyl]tetrahydropyran-3,4,5- J = 11.5, 5.8, 3.3 Hz, 2H), 3.63 triol(td, J = 9.5, 3.0 Hz, 2H), 2.15-2.07 (m, 2H), 1.99-1.89 (m, 4H).Preparation of Compound 200 (Method D)

tert-Butyl2,7-bis[2-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]ethynyl]spiro[fluorene-9,4′-piperidine]-1′-carboxylate

Compound 200 is prepared according to the procedure described forCompound 182 but using Intermediates AG5 (42.0 mg, 0.0850 mmol), M (400μL of 0.53 M, 0.2130 mmol). The reaction mixture is sealed and heated at100° C. for 2 h under N₂. Purification by reverse phase HPLC followed bylyophilisation afforded the title compound (11 mg, 18%) as a off-whitesolid. ¹H NMR (400 MHz, CD₃OD) δ 7.83-7.75 (m, 4H), 7.49 (d, J=7.8 Hz,2H), 4.02 (d, J=2.4 Hz, 2H), 3.95 (dd, J=9.4, 3.1 Hz, 2H), 3.92-3.79 (m,8H), 3.73 (dd, J=11.4, 5.5 Hz, 2H), 3.63 (t, J=9.4 Hz, 2H), 1.83 (s,4H), 1.52 (s, 9H). (Two protons of C-1 sugar are underneath of solventpeak). ESI-MS m/z calc. 707.2942. found 708.22 (M+1)+.

Preparation of Compound 201 (Method D)

(2R,3S,4R,5S,6R)-2-(hydroxymethyl)-6-[2-[F-methyl-7-[2-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]ethynyl]spiro[fluorene-9,4′-piperidine]-2-yl]ethynyl]tetrahydropyran-3,4,5-triol

Compound 201 is prepared according to the procedure described forCompound 182 but using Intermediates AG14. ¹H NMR (400 MHz, CD₃OD) δ7.88-7.80 (m, 4H), 7.54 (d, J=7.9 Hz, 2H), 4.04-4.00 (m, 2H), 3.93 (dd,J=9.4, 3.2 Hz, 2H), 3.91-3.80 (m, 4H), 3.74 (dd, J=11.4, 5.7 Hz, 2H),3.63 (t, J=9.4 Hz, 2H), 3.52-3.44 (m, 4H), 3.00 (s, 3H), 2.20-1.98 (m,4H). (two protons of C-1 sugar are underneath of solvent peak). ESI-MSm/z (M+H)⁺ 622.59

Preparation of Compound 202 (Method D)

1-[2,7-Bis[2-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]ethynyl]spiro[fluorene-9,4′-piperidine]-1′-yl]ethanone

A mixture of Intermediates M (340 μL of 0.53 M, 0.1802 mmol), AG8, (35mg, 0.080 mmol), Pd(dppf)Cl₂— CH₂Cl₂ (4.3 mg, 0.0053 mmol), CuI (4.6 mg,0.024 mmol) in DMF (140 μL) is degassed (vacuum/N₂). To this is addedDIPEA (112 μL, 0.6430 mmol), degassed. The reaction mixture is heated at100° C. for 2 h, additional amount of Intermediate M (152 μL of 0.53 M,0.080 mmol) is added and the resulting mixture is heated for anadditional 2.5 h under N₂. The reaction mixture cooled to RT, filteredthrough metal scavenger (Si-DMT; Silicycle; SPE-R79030B-06P) cartridge,washed with DMF (1 mL). The filtrate is directly purified by reversephase HPLC to afford the title compound (42 mg, 80%) as a white solid.¹H NMR (400 MHz, CD₃OD) δ 7.82-7.78 (m, 4H), 7.50 (d, J=8.0 Hz, 2H),4.05-3.98 (m, 4H), 3.98-3.91 (m, 4H), 3.91-3.79 (m, 4H), 3.73 (dd,J=11.4, 5.6 Hz, 2H), 3.63 (t, J=9.4 Hz, 2H), 2.25-2.19 (m, 3H),1.98-1.89 (m, 2H), 1.88-1.80 (m, 2H) (two protons of C-1 sugar areunderneath solvent peak). ESI-MS m/z calc. 649.2523. found 650.6 (M+1)⁺.

Alternative Preparation of Compound 202

1-[2,7-Bis[2-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]ethynyl]spiro[fluorene-9,4′-piperidine]-1′-yl]ethanone

Step I: Crude Compound 202

A mixture of Intermediates M (12.8 mL of 0.468 M, 5.99 mmol), AG8, (875mg, 1.999 mmol), Pd(dppf)Cl₂— CH₂Cl₂ (98 mg, 0.12 mmol), CuI (114 mg,0.6 mmol) in DMF (3.0 mL) is degassed (vacuum/N₂ flush twice 5 mineach). DIPEA (2.80 mL, 16.1 mmol) is added and the mixture is degassedtwice. The final reaction mixture is heated at 100° C. for 2 h (LC-MSafter 1 h showed complete conversion of AG8), cooled to RT andconcentrated under vacuo at 40° C. to afford the title compound as darkbrown oil.

Step II:[(2R,3R,4R,5R,6R)-3,4,5-triacetoxy-6-[2-[1′-acetyl-7-[2-[(2R,3R,4R,5R,6R)-3,4,5-triacetoxy-6-(acetoxymethyl)tetrahydropyran-2-yl]ethynyl]spiro[fluorene-9,4′-piperidine]-2-yl]ethynyl]tetrahydropyran-2-yl]methylAcetate

To a stirred solution of the crude Compound 202 from Step I in pyridine(9 mL) at RT is sequentially added DMAP (12 mg, 0.098 mmol) and aceticanhydride (3.8 mL, 40.3 mmol) at RT. The mixture is stirred for 16 h,diluted with water (40 mL), extracted with EtOAc (3×30 mL). The combinedextracts are washed with aqueous 1N HCl (3×20 mL), brine, passed throughphase separator and concentrated. The residue is purified on Biotage™SNAP silica gel cartridge (100 g) eluting with a gradient of EtOAC inHex (10% to 20%, 8 CV; and 100%) as eluent to afford the title compound(1.690 g, 86%) as a beige foam. To a stirred solution of the latter inEtOAc (10 mL) is added 300 mg of SiliaMetS Thiol (Cat # R51030B fromSilicycle) (1.42 mmol/g) and the mixture is stirred at RT for 3 h,filtered, washed with EtOAc (15 mL) and concentrated. The filtrate istreated one more time with SiliaMetS Thio to afford the title compound(1.600 g).

Step III: Compound 202

To a stirred solution of[(2R,3R,4R,5R,6R)-3,4,5-triacetoxy-6-[2-[1′-acetyl-7-[2-[(2R,3R,4R,5R,6R)-3,4,5-triacetoxy-6-(acetoxymethyl)tetrahydropyran-2-yl]ethynyl]spiro[fluorene-9,4′-piperidine]-2-yl]ethynyl]tetrahydropyran-2-yl]methylacetate (1.600 g) from Step II in MeOH (15 mL) is added MeONa in MeOH(400 μL of 0.5 M, 0.200 mmol). The mixture is stirred at RT for 7 h andquenched with AcOH (20 μL, 0.36 mmol). After stirring for 15 min, theresulting solid is filtered, washed with MeOH (10 mL), dried undervacuum oven at 40° C. for 40 h to afford the title compound (0.870 g,65% overall yield from Step I) as off-white solid.

Preparation of Compound 203 to 205

Compounds 203, 204 and 205 are prepared according to the proceduredescribed for Compound 182 but using the Intermediates AG42, AG43 andAG44 respectively.

LCMS m/z Compound IUPAC ¹H-NMR (M + H)⁺ 203 (2R, 3S, 4R, 5S, 6R)-2- (400MHz, CD₃OD) δ 7.92 (s, 623.29 (hydroxymethyl)-6-[2-[4- 1H), 7.76 (dd, J= 14.3, 7.9 Hz, hydroxy-7′-[2- 2H), 7.62 (s, 1H), 7.47 (ddd, J = [(2R,3S, 4R, 5S, 6R)-3,4,5- 18.4, 7.9, 1.3 Hz, 2H), 4.89 (d, trihydroxy-6- J= 2.1 Hz, 1H), 4.88 (d, 1H), (hydroxymethyl)tetrahydropyran- 4.11-3.91(m, 5H), 3.93-3.78 2-yl]ethynyl]spiro[cyclohexane- (m, 4H), 3.74 (ddd, J= 11.3, 1,9′-fluorene]-2′- 5.5, 1.5 Hz, 2H), 3.63 (td, J =yl]ethynyl]tetrahydropyran-3,4,5- 9.4, 1.5 Hz, 2H), 2.14-2.04 triol (m,2H), (2.02-1.92 m, 4H), 1.67 (m, 2H). 204 (2R, 3S, 4R, 5S, 6R)-2- (400MHz, CD₃OD) δ 7.82-7.65 637.3 (hydroxymethyl)-6-[2-[1- (m, 4H), 7.47(ddd, J = 10.5, 7.9, hydroxy-1-methyl-7′-[2- 1.4 Hz, 2H), 4.89 (m, 2H),4.02 [(2R, 3S, 4R, 5S, 6R)-3,4,5- (dd, J = 3.3, 2.1 Hz, 2H), 3.96trihydroxy-6- (ddd, J = 9.3, 4.4, 3.3 Hz, 2H),(hydroxymethyl)tetrahydropyran- 3.91-3.81 (m, 4H), 3.73 (dd, J =2-yl]ethynyl]spiro[cyclohexane- 11.4, 5.5 Hz, 2H), 3.63 (t, J = 9.44,9′-fluorene]-2′- Hz, 2H), 2.20-2.08 (m, 2H),yl]ethynyl]tetrahydropyran-3,4,5- 2.03-1.88 (m, 4H), 1.55-1.45 triol (m,2H), 1.44 (s,3H). 205 (2R, 3S, 4R, 5S, 6R)-2-[2-[4- (400 MHz, CDCl₃) δ8.05-7.98 622.28 amino-7′-[2-[(2R, 3S, 4R, 5S, (s, 1H), 7.83 (d, J = 7.9Hz, 1H), 6R)-3,4,5-trihydroxy-6- 7.76 (d, J = 7.9 Hz, 1H), 7.62 (s,(hydroxymethyl)tetrahydropyran- 1H), 7.53 (dd, J = 7.9, 1.2 Hz,2-yl]ethynyl]spiro[cyclohexane- 1H), 7.46 (dd, J = 7.9, 1.3 Hz,1,9′-fluorene]-2′-yl]ethynyl]-6- 1H), 4.88 (d, J = 2.2 Hz, 1H),(hydroxymethyl)tetrahydropyran- 4.86 (m, 1H), 4.01 (ddd, J = 7.3,3,4,5-triol 3.3, 2.1 Hz, 2H), 3.93 (ddd, J = 9.3, 3.3, 1.7 Hz, 2H),3.91-3.77 (m, 4H), 3.77-3.68 (m, 2H), 3.62 (td, J = 9.4, 4.7 Hz, 2H),3.48 (dd, J = 10.3, 5.1 Hz, 1H), 2.30-2.04 (m, 6H), 1.56 (d, J = 13.1Hz, 2H).Preparation of Compound 206 (Method D)

1-[2,7-Bis[2-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]ethynyl]spiro[fluorene-9,4′-piperidine]-1′-yl]-2-hydroxy-2-methyl-propan-1-one

Compound 206 is prepared according to the procedure described forCompound 182 but using Intermediates M (275 μL of 0.53 M, 0.15 mmol) andAG9, (34 mg, 0.0622 mmol). The reaction mixture is heated at 100° C. for2 h under N₂ and an additional amount of Intermediate M (120 μL of 0.53M, 0.064 mmol) is added to complete the reaction. The reaction mixtureis stirred at 100° C. for 2 h. Purification by reverse phase HPLCafforded the title compound (11 mg, 23%). ¹H NMR (400 MHz, CD₃OD) δ7.83-7.78 (m, 4H), 7.50 (dd, J=7.8, 1.3 Hz, 2H), 4.03-4.00 (m, 2H), 3.95(dd, J=9.3, 3.3 Hz, 2H), 3.90-3.80 (m, 4H), 3.73 (dd, J=11.4, 5.6 Hz,2H), 3.63 (t, J=9.4 Hz, 2H), 1.95-1.82 (m, 4H), 1.51 (s, 6H). Sixprotons are underneath solvent peaks. ESI-MS m/z calc. 693.2785. found694.58 (M+1)⁺.

Preparation of Compound 207 (Method D)

Methyl2,7-bis[2-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]ethynyl]spiro[fluorene-9,4′-piperidine]-1′-carboxylate

Compound 207 is prepared according to the procedure described forCompound 182 but using Intermediates M (375 μL of 0.53 M, 0.199 mmol)and AG10, (40 mg, 0.08405 mmol). The reaction mixture is heated at 100°C. for 2 h under N₂ and an additional amount of Intermediate M (160 μLof 0.53 M, 0.08480 mmol) is added to complete the reaction. The reactionmixture is stirred at 100° C. for 2 h. Purification by reverse phaseHPLC afforded the title compound (31 mg, 54%). ¹H NMR (400 MHz, CD₃OD) δ7.82-7.77 (m, 4H), 7.49 (dd, J=7.9, 1.0 Hz, 2H), 4.03-4.00 (m, 2H), 3.95(dd, J=9.4, 3.3 Hz, 2H), 3.93-3.80 (m, 8H), 3.76 (s, 3H), 3.73 (dd,J=11.4, 5.6 Hz, 2H), 3.63 (t, J=9.4 Hz, 2H), 1.89-1.79 (m, 4H). Twoprotons for C-1 sugar are under solvent peak. ESI-MS m/z calc.665.24725, found 666.57 (M+1)⁺.

Preparation of Compound 208 and 209

Compounds 208 and 209 are prepared according to the procedure describedfor Compound 207 but using the appropriate Intermediates prepared asdescribed for Intermediates AG10

LCMS m/z Compound IUPAC name ¹H-NMR (M + H)⁺ 208 isopropyl 2,7-bis[2-(400 MHz, CD₃OD) δ 694.27 [(2R, 3S, 4R, 5S, 6R)-3,4,5- 7.81-7.75 (m,4H), 7.49 trihydroxy-6- (dd, J = 7.9, 1.3 Hz, 2H),(hydroxymethyl)tetrahydropyran- 4.99-4.90 (m, 1H), 4.88 (d,2-yl]ethynyl]spiro[fluorene-9,4′- J = 2.1 Hz, 2H), 4.02 (dd,piperidine]-1′-carboxylate J = 3.2, 2.2 Hz, 2H), 3.95 (dd, J = 9.3, 3.3Hz, 2H), 3.92-3.80 (m, 8H), 3.73 (dd, J = 11.4, 5.6 Hz, 2H), 3.63 (t, J= 9.4 Hz, 2H), 1.88-1.77 (m, 4H), 1.30 (d, J = 6.2 Hz, 6H). 209 ethyl2,7-bis[2- (400 MHz, CD₃OD) δ 680.26 [(2R, 3S, 4R, 5S, 6R)-3,4,5-7.82-7.74 (m, 4H), 7.49 trihydroxy-6- (dd, J = 7.8, 1.3 Hz, 2H),(hydroxymethyl)tetrahydropyran- 4.88 (d, J = 2.2 Hz, 2H),2-yl]ethynyl]spiro[fluorene-9,4′- 4.19 (q, J = 7.1 Hz, 2H),piperidine]-1′-carboxylate 4.02 (dd, J = 3.2, 2.2 Hz, 2H), 3.95 (dd, J =9.3, 3.3 Hz, 2H), 3.93-3.80 (m, 8H), 3.73 (dd, J = 11.4, 5.6 Hz, 2H),3.63 (t, J = 9.4 Hz, 2H), 1.87-1.78 (m, 4H), 1.31 (t, J = 7.1 Hz, 3H).Preparation of Compound 210 (Method D)

(2R,3S,4R,5S,6R)-2-(hydroxymethyl)-6-[2-[1′-methylsulfonyl-7-[2-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]ethynyl]spiro[fluorene-9,4′-piperidine]-2-yl]ethynyl]tetrahydropyran-3,4,5-triol

Compound 210 is prepared according to the procedure described forCompound 182 but using Intermediates M (350 μL of 0.53 M, 0.186 mmol)and AG11, (40 mg, 0.08 mmol). The reaction mixture is stirred at 100° C.for 2 h under N₂ then an additional amount of Intermediate M (150 μL of0.53 M, 0.08 mmol) is added and the final mixture is stirred at 100° C.for 2 h. Purification by reverse phase HPLC afforded the title compound(23 mg, 41%). ¹H NMR (400 MHz, CD₃OD) δ 7.85-7.77 (m, 4H), 7.51 (d,J=9.1 Hz, 2H), 4.02 (dd, J=3.2, 2.2 Hz, 2H), 3.95 (dd, J=9.3, 3.3 Hz,2H), 3.90-3.81 (m, 4H), 3.76-3.66 (m, 6H), 3.63 (t, J=9.4 Hz, 2H), 3.06(s, 3H), 2.03-1.95 (m, 4H). Two protons of C-1 sugar are under thesolvent peak. ESI-MS m/z calc. 685.2193. found 686.55 (M+1)⁺.

Preparation of Compound 211 (Method D)

[9-(acetoxymethyl)-2,7-bis[2-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]ethynyl]fluoren-9-yl]methylAcetate

Compound 211 is prepared according to the procedure described forCompound 182 but using Intermediates M (1 mL of 0.53 M, 0.53 mmol) andAG12 (80 mg, 0.17 mmol). Purification by reverse phase HPLC afforded thetitle compound (36 mg, 29%). ¹H NMR (400 MHz, CD₃OD) δ 7.82 (d, J=7.9Hz, 2H), 7.71 (s, 2H), 7.55 (d, J=7.9 Hz, 2H), 4.88 (d, J=2.1 Hz, 2H),4.40 (s, 4H), 4.02 (dd, J=3.2, 2.2 Hz, 2H), 3.95 (dd, J=9.3, 3.3 Hz,2H), 3.90-3.80 (m, 4H), 3.73 (dd, J=11.4, 5.5 Hz, 2H), 3.63 (t, J=9.4Hz, 2H), 1.98 (s, 6H). ESI-MS m/z calc. 682.22614. found 683.22 (M+1)⁺.

Preparation of Compound 212 (Method D)

(2R,3S,4R,5S,6R)-2-(hydroxymethyl)-6-[2-[7-[2-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]ethynyl]-1H-indazol-5-yl]ethynyl]tetrahydropyran-3,4,5-triol

To a reaction tube charged with 5-bromo-7-iodo-1H-indazole (45.0 mg,0.139 mmol) prepared following the procedure described in PCT Int.Appl., 2007117465, Pd(dppf)Cl₂. CH₂Cl₂ (6.0 mg, 0.0082 mmol) and CuI(6.0 mg, 0.032 mmol), capped and degassed (vacuum then nitrogen flush,2×) is added Intermediate M (500 μL of 0.53 M, 0.265 mmol) as a solutionin DMF and DIPEA (400 μL). The reaction tube is degassed again,transferred to a preheated (80° C.) oil bath and stirred overnight.After cooling down to RT, the reaction mixture is passed through a 200mg Si-DMT cartridge, rinsed with portions of MeOH and purified byreverse phase HPLC. The fractions are combined and freeze-dried,providing the title compound (18.2 mg, 27% yield) as a fluffy whitesolid. ¹H NMR (400 MHz, CD3OD) δ 8.14 (s, 1H), 7.97 (d, J=1.3 Hz, 1H),7.60 (d, J=1.2 Hz, 1H), 5.01 (d, J=2.1 Hz, 1H), 4.93-4.78 (m, 1H),4.16-4.09 (m, 1H), 4.06-4.01 (m, 1H), 4.01-3.81 (m, 6H), 3.80-3.70 (m,2H), 3.69-3.58 (m, 2H). ESI-MS m/z: 491.44 (M+1)⁺

Preparation of Compounds 213 to 216.

Compounds 213-216 are prepared according to the procedure described forCompound 182 but using the Intermediates AG30 to AG33 respectively

LCMS m/z Compound IUPAC name ¹H-NMR (M + H)⁺ 213 (2R, 3S, 4R, 5S,6R)-2-[2-[9-[2- (400 MHz, CD₃OD) δ 7.79 (d, J = 639.31(dimethylamino)ethyl-methyl- 7.6 Hz, 4H), 7.53 (d, J = 7.8 Hz,amino]-7-[2-[(2R, 3S, 4R, 5S, 2H), 5.03 (s, 1H), 4.87 (s, 2H),6R)-3,4,5-trihydroxy-6- 4.00 (t, J = 2.7 Hz, 2H), 3.95-3.88(hydroxymethyl)tetrahydropyran- (m, 3H), 3.86 (d, J = 2.2 Hz,2-yl]ethynyl]-9H-fluoren-2- 1H), 3.85-3.79 (m, 2H), 3.73 yl]ethynyl]-6-(ddd, J = 11.4, 5.9, 1.4 Hz, 2H), (hydroxymethyl)tetrahydropyran- 3.63(td, J = 9.5, 1.0 Hz, 2H), 3,4,5-triol 3.23-3.17 (m, 2H), 2.81 (s, 6H),2.34 (s, 3H). 214 (2R, 3S, 4R, 5S, 6R)-2-[2-[9- (400 MHz, CD₃OD) δ 7.67(d, J = 637.27 (cyclopentylmethyl)-9-hydroxy- 7.8 Hz, 2H), 7.58-7.53 (m,2H), 7-[2-[(2R, 3S, 4R, 5S, 6R)-3,4,5- 7.47 (dd, J = 7.8, 1.4 Hz, 2H),trihydroxy-6- 4.88 (d, J = 2.4 Hz, 2H), 4.02 (ddt,(hydroxymethyl)tetrahydropyran- J = 3.0, 2.0, 1.0 Hz, 2H), 3.95 (dt,2-yl]ethynyl]fluoren-2- J = 9.3, 3.2 Hz, 2H), 3.91-3.88 yl]ethynyl]-6-(m, 1H), 3.87-3.80 (m, 3H), 3.73 (hydroxymethyl)tetrahydropyran- (dd, J= 11.2, 5.6 Hz, 2H), 3.63 (t, 3,4,5-triol J = 9.5 Hz, 2H), 2.29 (d, J =6.2 Hz, 2H), 1.43-1.33 (m, 2H), 1.21-1.04 (m, 4H), 1.03-0.92 (m, 1H),0.82-0.70 (m, 2H). 215 (2R, 3S, 4R, 5S, 6R)-2-[2-[9- (400 MHz, CD₃OD) δ7.68 (d, J = 651.27 (cyclohexylmethyl)-9-hydroxy-7- 7.8 Hz, 2H), 7.55(s, 2H), 7.47 [2-[(2R, 3S, 4R, 5S, 6R)-3,4,5- (dd, J = 7.8, 1.3 Hz, 2H),4.89 (d, trihydroxy-6- J = 2.2 Hz, 2H), 4.06-4.00 (m,(hydroxymethyl)tetrahydropyran- 2H), 3.95 (dt, J = 9.3, 3.5 Hz, 2H),2-yl]ethynyl]fluoren-2- 3.91-3.80 (m, 4H), 3.73 (dd, J = yl]ethynyl]-6-11.3, 5.5 Hz, 2H), 3.63 (t, J = 9.4 (hydroxymethyl)tetrahydropyran- Hz,2H), 2.15-2.07 (m, 2H), 3,4,5-triol 1.46-1.36 (m, 3H), 1.11 (d, J = 10.3Hz, 2H), 1.04-0.90 (m, 1H), 0.90-0.80 (m, 2H), 0.74 (td, J = 15.7, 12.7,5.1 Hz, 2H), 0.69-0.58 (m, 1H). 216 (2R, 3S, 4R, 5S, 6R)-2-[2-[9- (400MHz, CD₃OD) δ 7.68 (d, J = 599.27 hydroxy-9-(2-hydroxyethyl)-7- 7.9 Hz,2H), 7.62-7.59 (m, 2H), [2-[(2R, 3S, 4R, 5S, 6R)-3,4,5- 7.48 (dd, J =7.9, 1.4 Hz, 2H), trihydroxy-6- 4.88 (d, J = 1.1 Hz, 2H), 4.03-4.00(hydroxymethyl)tetrahydropyran- (m, 2H), 3.96-3.92 (m, 2H),2-yl]ethynyl]fluoren-2- 3.90-3.79 (m, 4H), 3.73 (dd, J = yl]ethynyl]-6-11.5, 5.6 Hz, 2H), 3.63 (t, J = 9.5 (hydroxymethyl)tetrahydropyran- Hz,2H), 3.17-3.08 (m, 2H), 3,4,5-triol 2.41-2.32 (m, 2H).Preparation of Compound 217

(2R,3S,4R,5S,6R)-2-[2-[9,9-Bis(hydroxymethyl)-7-[2-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]ethynyl]fluoren-2-yl]ethynyl]-6-(hydroxymethyl)tetrahydropyran-3,4,5-triol

To a stirred solution of Compound 214 (23 mg, 0.03201 mmol) in MeOH (1mL) is added MeONa in MeOH (100 μL of 0.5 M, 0.05 mmol), reactionmixture is stirred at RT overnight, quenched with DOWEX 50WX4 hydrogenform resin until pH 4-5, filtered and concentrated to afford the titlecompound (15 mg, 68%) as off-white solid. ¹H NMR (400 MHz, CD₃OD) δ 7.77(d, J=7.9 Hz, 2H), 7.72 (s, 2H), 7.49 (dd, J=7.9, 1.3 Hz, 2H), 4.03-3.99(m, 2H), 3.95 (dd, J=9.3, 3.3 Hz, 2H), 3.90-3.80 (m, 8H), 3.73 (dd,J=11.3, 5.4 Hz, 2H), 3.63 (t, J=9.4 Hz, 2H), two protons for Cl—H sugaris under the solvent peak. ESI-MS m/z calc. 598.205. found 599.53(M+1)⁺.

Preparation of Compound 218 (Method D)

2,7-bis[2-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]ethynyl]spiro[fluorene-9,5′-oxepane]-2′-one

Compound 218 is prepared according to the procedure described forCompound 182 but using the Intermediates AG13. ¹H NMR (400 MHz, CD₃OD) δ7.90 (s, 1H), 7.84-7.75 (m, 2H), 7.60-7.46 (m, 3H), 4.81-4.70 (m, 1H),4.04-4.00 (m, 2H), 3.99-3.93 (m, 2H), 3.91-3.81 (m, 4H), 3.73 (dd,J=11.2, 5.4 Hz, 2H), 3.63 (t, J=9.4 Hz, 2H), 3.21-3.12 (m, 1H),2.85-2.74 (m, 1H), 2.44-1.94 (m, 4H), 1.44-1.34 (m, 1H). C-1 sugarprotons are under the solvent peak. ESI-MS m/z calc. 637.27 (M+H)⁺.

Preparation of Compound 219

3-[9-(2-hydroxyethyl)-2,7-bis[2-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]ethynyl]fluoren-9-yl]propanoicAcid

To a suspension of Compound 218 (0.075 mmol) in 1,4-dioxane (0.5 mL) isadded aqueous LiOH in Water (100 μL of 1 M, 0.100 mmol). The resultingreaction mixture is stirred at RT for 5 h, quenched with AcOH (20 μL),purified by reverse phase HPLC to afford the title compound (7 mg, 14%)as white solid. ¹H NMR (400 MHz, CD₃OD) δ 7.77 (d, J=7.9 Hz, 2H), 7.58(s, 2H), 7.49 (d, J=7.9 Hz, 2H), 4.04-4.00 (m, 2H), 3.96 (dd, J=9.3, 3.1Hz, 2H), 3.91-3.81 (m, 4H), 3.73 (dd, J=11.3, 5.4 Hz, 2H), 3.63 (t,J=9.4 Hz, 2H), 2.84-2.73 (m, 2H), 2.46-2.29 (m, 4H), 1.48-1.34 (m, 2H).C-1 Sugar protons are under the solvent peak. ESI-MS m/z calc. 655.21(M+H)⁺.

Preparation of Compound 220 (Method D)

[2,7-Bis[2-[(2R,3S,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]ethynyl]spiro[fluorene-9,4′-piperidine]-1′-yl]-cyclopropyl-methanone

Step I: Crude Compound 192

A mixture of Intermediates AG4, (84 mg, 0.186 mmol), M (1.1 mL of 0.53M, 0.583 mmol), Pd(dppf)Cl₂— CH₂Cl₂ (10.0 mg, 0.0123 mmol), CuI (11 mg,0.058 mmol) in DMF (320 μL) is degassed (vacuum/N₂). To the reactionmixture is added DIPEA (260 μL, 1.493 mmol), degassed. The reactionmixture is placed on preheated oil bath at 100° C., stirred for 2 hunder N₂, cooled to RT to afford the title compound. This crude reactionmixture is used as such in the next step without further workup.

Step II: Compound 220

To ⅓ of the crude reaction mixture from Step I (0.062 mmol, 0.55 mL) isadded to a mixture of HATU (25.5 mg, 0.067 mmol) andcyclopropanecarboxylic acid (5.8 mg, 0.067 mmol) at RT. The reactionmixture is stirred for 2 h. The crude reaction mixture is filteredthrough metal scavenger (Si-DMT; Silicycle; SPE-R79030B-06P), washedwith 0.5 mL of DMF. The filtrate is directly purified by reverse phaseHPLC to afford the title compound (18 mg, 50%). ¹H NMR (400 MHz, CD₃OD)δ 7.83-7.77 (m, 4H), 7.50 (dd, J=8.0, 1.0 Hz, 2H), 4.88 (d, J=2.1 Hz,2H), 4.24-4.15 (m, 2H), 4.06-3.99 (m, 4H), 3.95 (dd, J=9.3, 3.3 Hz, 2H),3.90-3.80 (m, 4H), 3.73 (dd, J=11.4, 5.6 Hz, 2H), 3.63 (t, J=9.4 Hz,2H), 2.14-2.04 (m, 1H), 1.99-1.90 (m, 2H), 1.89-1.80 (m, 2H), 1.00-0.94(m, 2H), 0.90-0.82 (m, 2H). ESI-MS m/z calc. 675.26794. found 676.51(M+1)⁺.

Preparation of Compounds 221 to 245.

Compounds 221 to 245 are prepared according to the procedure describedfor Compound 220 but using the appropriate commercially availablecarboxylic acid.

LCMS m/z Compound IUPAC name ¹H NMR (M + H)⁺ 221 1-[2,7-bis[2-[(2R, 3S,4R, 5S, (400 MHz, CD₃OD) δ 8.27 (s, 735.62 6R)-3,4,5-trihydroxy-6- 1H),7.84-7.77 (m, 4H), 7.51 (hydroxymethyl)tetrahydropyran- (d, J = 7.9 Hz,2H), 4.10-3.43 2-yl]ethynyl]spiro[fluorene-9,4′- (m, 22H), 2.82-2.65 (m,4H), piperidine]-1′-yl]-2-morpholino- 1.99-1.80 (m, 4H). [1] ethanone222 1-[2,7-bis[2-[(2R, 3S, 4R, 5S, (400 MHz, CD₃OD) δ 8.45 (s, 693.476R)-3,4,5-trihydroxy-6- 1H), 7.81-7.74 (m, 4H), 7.50(hydroxymethyl)tetrahydropyran- (d, J = 7.9 Hz, 2H), 4.22 (s,2-yl]ethynyl]spiro[fluorene-9,4′- 2H), 4.07-3.99 (m, 4H), 3.93piperidine]-1′-yl]-2- (dd, J = 9.3, 3.2 Hz, 2H), (dimethylamino)ethanone3.87-3.79 (m, 4H), 3.73 (dd, J = 11.4, 5.6 Hz, 2H), 3.63 (t, J = 9.4 Hz,2H), 2.91 (s, 6H), 1.98-1.80 (m, 4H). 223 1-[2,7-bis[2-[(2R, 3S, 4R, 5S,(400 MHz, CD₃OD) δ 7.82-7.77 664.18 6R)-3,4,5-trihydroxy-6- (m, 4H),7.50 (d, J = 8.0 (hydroxymethyl)tetrahydropyran- Hz, 2H), 4.05-3.98 (m,4H), 2-yl]ethynyl]spiro[fluorene-9,4′- 3.98-3.92 (m, 4H), 3.90-3.80piperidine]-1′-yl]propan-1-one (m, 4H), 3.73 (dd, J = 11.4, 5.6 Hz, 2H),3.63 (t, J = 9.4 Hz, 2H), 2.54 (q, J = 7.4 Hz, 2H), 1.95-1.79 (m, 4H),1.19 (t, J = 7.5 Hz, 3H). C-1 sugar protons are under the solvent peak.224 1-[2,7-bis[2-[(2R, 3S, 4R, 5S, (400 MHz, CD₃OD) δ 7.83-7.78 678.186R)-3,4,5-trihydroxy-6- (m, 4H), 7.50 (d, J = 8.0(hydroxymethyl)tetrahydropyran- Hz, 2H), 4.07-3.97 (m, 6H),2-yl]ethynyl]spiro[fluorene-9,4′- 3.95 (dd, J = 9.4, 3.3 Hz, 2H),piperidine]-1′-yl]-2-methyl- 3.90-3.80 (m, 4H), 3.73 (dd, propan-1-one J= 11.4, 5.6 Hz, 2H), 3.63 (t, J = 9.4 Hz, 2H), 3.14-3.02 (m, 1H),1.95-1.81 (m, 4H), 1.19 (d, J = 6.7 Hz, 6H). C-1 sugar protons are underthe solvent peak. 225 [2,7-bis[2-[(2R, 3S, 4R, 5S, 6R)- (400 MHz, CD₃OD)δ 7.86-7.76 690.21 3,4,5-trihydroxy-6- (m, 4H), 7.50 (d, J = 7.8(hydroxymethyl)tetrahydropyran- Hz, 2H), 4.18-3.99 (m, 6H),2-yl]ethynyl]spiro[fluorene-9,4′- 3.99-3.91 (m, 2H), 3.91-3.78piperidine]-1′-yl]-(l- (m, 4H), 3.73 (dd, J = 11.3,methylcyclopropyl)methanone 5.5 Hz, 2H), 3.63 (t, J = 9.4 Hz, 2H),1.96-1.79 (m, 4H), 1.40 (s, 3H), 1.08-0.98 (m, 2H), 0.74-0.60 (m, 2H).C-1 sugar protons are under the solvent peak. 226 [2,7-bis[2-[(2R, 3S,4R, 5S, 6R)- (400 MHz, CD₃OD) δ 7.83-7.75 690.21 3,4,5-trihydroxy-6-7.75 (m, 4H), 7.50 (d, J = 7.9 (hydroxymethyl)tetrahydropyran- Hz, 2H),4.04-3.97 (m, 4H), 2-yl]ethynyl]spiro[fluorene-9,4′- 3.94 (dd, J = 9.3,3.3 Hz, 2H), piperidine]-1′-yl]-cyclobutyl- 3.90-3.80 (m, 6H), 3.73 (dd,methanone J = 11.4, 5.6 Hz, 2H), 3.63 (t, J = 9.4 Hz, 2H), 3.58-3.46 (m,1H), 2.45-2.32 (m, 2H), 2.29-2.19 (m, 2H), 2.11-1.97 (m, 1H), 1.93-1.78(m, 5H). C-1 sugar protons are under the solvent peak. 227[2,7-bis[2-[(2R, 3S, 4R, 5S, 6R)- (400 MHz, CD₃OD) δ 7.85-7.76 692.193,4,5-trihydroxy-6- (m, 4H), 7.50 (d, J = 7.9(hydroxymethyl)tetrahydropyran- Hz, 2H), 4.42-3.9 (br m, 4H),2-yl]ethynyl]spiro[fluorene-9,4′- 4.06-3.99 (m, 2H), 3.95 (dd,piperidine]-1′-yl]-(1- J = 9.4, 3.2 Hz, 2H), 3.91-3.80hydroxycyclopropyl)methanone (m, 4H), 3.78-3.70 (m, 2H), 3.63 (t, J =9.4 Hz, 2H), 1.99-1.83 (m, 4H), 1.19-1.10 (m, 2H), 0.98-0.89 (m, 2H).C-1 sugar protons are under the solvent peak. 228 1-[2,7-bis[2-[(2R, 3S,4R, 5S, (400 MHz, CD₃OD) δ 692.22 6R)-3,4,5-trihydroxy-6- 7.83-7.78 (m,4H), (hydroxymethyl)tetrahydropyran- 7.53-7.47 (m, 2H), 4.13-4.052-yl]ethynyl]spiro[fluorene-9,4′- (m, 4H), 4.04-4.00 (m, 2H),piperidine]-1′-yl]-2,2-dimethyl- 3.95 (dd, J = 9.3, 3.3 Hz, 2H),propan-1-one 3.91-3.80 (m, 4H), 3.73 (dd, J = 11.4, 5.6 Hz, 2H), 3.63(t, J = 9.4 Hz, 2H), 1.93-1.81 (m, 4H), 1.37 (s, 9H). C-1 sugar protonsare under the solvent peak. 229 (2R)-1-[2,7-bis[2- (400 MHz, CD₃OD) δ7.84-7.76 694.21 [(2R, 3S, 4R, 5S, 6R)-3,4,5- (m, 4H), 7.51 (d, J = 7.8trihydroxy-6- Hz, 2H), 4.39 (q, J = 6.6 Hz,(hydroxymethyl)tetrahydropyran- 1H), 4.11-4.00 (m, 6H), 3.942-yl]ethynyl]spiro[fluorene-9,4′- (dd, J = 9.4, 3.3 Hz, 2H),piperidine]-1′-yl]-2-methoxy- 3.90-3.79 (m, 4H), 3.73 (dd, propan-1-oneJ = 11.4, 5.6 Hz, 2H), 3.63 (t, J = 9.4 Hz, 2H), 3.40 (s, 3H), 1.98-1.81(m, 4H), 1.42 (d, J = 6.7 Hz, 3H). C-1 sugar protons are under thesolvent peak. 230 1-[2,7-bis[2-[(2R, 3S, 4R, 5S, (400 MHz, CD₃OD) δ 7.84(s, 701.16 6R)-3,4,5-trihydroxy-6- 2H), 7.81 (d, J = 7.9 Hz, 2H),(hydroxymethyl)tetrahydropyran- 7.52 (d, J = 7.9 Hz, 2H),2-yl]ethynyl]spiro[fluorene-9,4′- 4.36-3.99 (m, 6H), 3.95 (dd,piperidine]-1′- J = 9.3, 3.2 Hz, 2H), 3.90-3.80carbonyl]cyclopropanecarbonitrile (m, 4H), 3.73 (dd, J = 11.3, 5.5 Hz,2H), 3.63 (t, J = 9.3 Hz, 2H), 2.08-1.81 (m, 4H), 1.73-1.58 (m, 4H). C-1sugar protons are under the solvent peak. 231 [2,7-bis[2-[(2R, 3S, 4R,5S, 6R)- (400 MHz, CD₃OD) δ 7.85 (s, 702.17 3,4,5-trihydroxy-6- 2H),7.80 (d, J = 7.9 Hz, 2H), (hydroxymethyl)tetrahydropyran- 7.77-7.72 (m,1H), 7.50 (d, 2-yl]ethynyl]spiro[fluorene-9,4′- J = 7.9 Hz, 2H), 6.70(d, J = 2.4 piperidine]-1′-yl]-(1H-pyrazol-5- Hz, 1H), 4.35-4.14 (m,4H), yl)methanone 4.05-4.00 (m, 2H), 3.96 (dd, J = 9.3, 3.3 Hz, 2H),3.91-3.80 (m, 4H), 3.73 (dd, J = 11.3, 5.4 Hz, 2H), 3.63 (t, J = 9.4 Hz,2H), 2.01-1.88 (m, 4H). C-1 sugar protons are under the solvent peak.232 [2,7-bis[2-[(2R, 3S, 4R, 5S, 6R)- (400 MHz, CD₃OD) δ 8.53 (d, 703.153,4,5-trihydroxy-6- 1H), 7.87 (s, 2H), 7.81 (d, J =(hydroxymethyl)tetrahydropyran- 7.9 Hz, 2H), 7.51 (d, J = 7.82-yl]ethynyl]spiro[fluorene-9,4′- Hz, 2H), 6.92 (d, 1H),piperidine]-1′-yl]-isoxazol-5-yl- 4.24-4.16 (m, 2H), 4.12-4.05 methanone(m, 2H), 4.04-4.00 (m, 2H), 3.98-3.92 (m, 2H), 3.90-3.81 (m, 4H),3.78-3.69 (m, 2H), 3.63 (t, J = 9.3 Hz, 2H), 2.03-1.93 (m, 4H). C-1sugar protons are under the solvent peak. 233 [2,7-bis[2-[(2R, 3S, 4R,5S, 6R)- (400 MHz, CD₃OD) δ 706.2 3,4,5-trihydroxy-6- 7.86-7.76 (m, 4H),7.54-7.48 (hydroxymethyl)tetrahydropyran- (m, 2H), 4.15-3.79 (m, 15H),2-yl]ethynyl]spiro[fluorene-9,4′- 3.73 (dd, J = 11.3, 5.6 Hz,piperidine]-1′-yl]-[(2R)- 2H), 3.63 (t, J = 9.4 Hz,tetrahydrofuran-2-yl]methanone 2H), 2.32-1.71 (m, 8H). C-1 sugar protonsare under the solvent peak. 234 4-[2,7-bis[2-[(2R, 3S, 4R, 5S, (400 MHz,CD₃OD) δ 7.92 (d, 707.18 6R)-3,4,5-trihydroxy-6- J = 7.9 Hz, 2H), 7.83(s, 2H), (hydroxymethyl)tetrahydropyran- 7.48 (d, J = 7.8 Hz, 2H), 7.312-yl]ethynyl]spiro[fluorene-9,4′- (s, 1H), 6.74 (s, 1H), 4.95 (d,piperidine]-1′-yl]-4-oxo- J = 4.3 Hz, 2H), 4.78 (d, J = butanamide 5.8Hz, 2H), 4.75-4.67 (m, 4H), 4.47 (t, J = 5.9 Hz, 2H), 3.89-3.33 (m,16H), 2.62 (t, J = 7.1 Hz, 2H), 2.36 (t, J = 7.0 Hz, 2H), 1.90-1.79 (m,2H), 1.77-1.65 (m, 2H). 235 [2,7-bis[2-[(2R, 3S, 4R, 5S, 6R)- (400 MHz,CD₃OD) δ 7.84 (s, 712.18 3,4,5-trihydroxy-6- 1H), 7.81 (d, J = 8.0 Hz,2H), (hydroxymethyl)tetrahydropyran- 7.73 (s, 1H), 7.51 (d, J = 7.92-yl]ethynyl]spiro[fluorene-9,4′- Hz, 2H), 4.16-3.99 (m, 6H),piperidine]-1′-yl]-(2,2- 3.95 (dd, J = 9.3, 3.1 Hz, 2H),difluorocyclopropyl)methanone 3.89-3.80 (m, 4H), 3.73 (dd, J = 11.2, 5.2Hz, 2H), 3.63 (t, J = 9.3 Hz, 2H), 3.13-3.02 (m, 1H), 2.13-1.78 (m, 6H).C-1 sugar protons are under the solvent peak. 236 [2,7-bis[2-[(2R, 3S,4R, 5S, 6R)- (400 MHz, CD₃OD) δ 8.96 (s, 714.16 3,4,5-trihydroxy-6- 1H),8.71 (d, J = 2.5 Hz, 1H), (hydroxymethyl)tetrahydropyran- 8.68 (d, J =1.5 Hz, 1H), 7.88 2-yl]ethynyl]spiro[fluorene-9,4′- (s, 2H), 7.81 (d, J= 7.9 Hz, piperidine]-1′-yl]-pyrazin-2-yl- 2H), 7.51 (d, J = 7.8 Hz,2H), methanone 4.28-4.19 (m, 2H), 4.05-4.01 (m, 2H), 3.99-3.93 (m, 4H),3.91-3.80 (m, 4H), 3.77-3.70 (m, 2H), 3.63 (t, J = 9.2 Hz, 2H),2.06-1.88 (m, 4H). C-1 sugar protons are under the solvent peak 237[2,7-bis[2-[(2R, 3S, 4R, 5S, 6R)- (400 MHz, CD₃OD) δ 7.88 (s, 716.183,4,5-trihydroxy-6- 2H), 7.81 (d, J = 7.9 Hz, 2H),(hydroxymethyl)tetrahydropyran- 7.55-7.48 (m, 3H), 6.63 (d,2-yl]ethynyl]spiro[fluorene-9,4′- J = 1.9 Hz, 1H), 4.26-4.14 (m,piperidine]-1′-yl]-(2- 2H), 4.05-4.00 (m, 4H), 4.00methylpyrazol-3-yl)methanone (s, 3H), 3.95 (dd, J = 9.3, 3.2 Hz, 2H),3.91-3.81 (m, 4H), 3.73 (dd, J = 11.3, 5.4 Hz, 2H), 3.63 (t, J = 9.3 Hz,2H), 2.03-1.82 (m, 4H). C-1 sugar protons are under the solvent peak.238 [2,7-bis[2-[(2R, 3S, 4R, 5S, 6R)- (400 MHz, CD₃OD) δ 7.85 (s, 716.213,4,5-trihydroxy-6- 2H), 7.81 (d, J = 7.9 Hz, 2H),(hydroxymethyl)tetrahydropyran- 7.67 (brs, 1H), 7.51 (d, J = 7.92-yl]ethynyl]spiro[fluorene-9,4′- Hz, 2H), 6.67 (brs, 1H),piperidine]-1′-yl]-(1 - 4.38-4.29 (m, 2H), 4.23-4.14methylpyrazol-3-yl)methanone (m, 2H), 4.02 (dd, J = 3.2, 2.1 Hz, 2H),3.98-3.92 (m, 5H), 3.90-3.81 (m, 4H), 3.73 (dd, J = 11.4, 5.6 Hz, 2H),3.63 (t, J = 9.4 Hz, 2H), 2.00-1.88 (m, 4H). C-1 sugar protons are underthe solvent peak. 239 [2,7-bis[2-[(2R, 3S, 4R, 5S, 6R)- (400 MHz, CD₃OD)δ 7.92-7.83 727.2 3,4,5-trihydroxy-6- (m, 3H), 7.80 (d, J = 7.9(hydroxymethyl)tetrahydropyran- Hz, 2H), 7.53-7.46 (m, 3H),2-yl]ethynyl]spiro[fluorene-9,4′- 7.39 (d, J = 7.8 Hz, 1H), 4.89piperidine]-1′-yl]-(6-methyl-2- (d, J = 1.9 Hz, 2H), 4.25-4.17pyridyl)methanone (m, 2H), 4.05-4.01 (m, 2H), 3.96 (dd, J = 9.3, 3.1 Hz,2H), 3.92-3.71 (m, 8H), 3.64 (t, J = 9.4 Hz, 2H), 2.59 (s, 3H),2.03-1.96 (m, 2H), 1.93-1.84 (m, 2H). 240 1-[2,7-bis[2-[(2R, 3S, 4R, 5S,(400 MHz, CD₃OD) δ 7.79 (d, 730.22 6R)-3,4,5-trihydroxy-6- J = 7.9 Hz,2H), 7.74 (s, 2H), (hydroxymethyl)tetrahydropyran- 7.71 (d, J = 2.3 Hz,1H), 7.55 2-yl]ethynyl]spiro[fluorene-9,4′- (d, J = 1.9 Hz, 1H), 7.49(dd, piperidine]-1′-yl]-3-pyrazol-1-yl- J = 7.9, 1.1 Hz, 2H), 6.34 (t, J= propan-1-one 2.1 Hz, 1H), 4.88 (d, J = 2.1 Hz, 3H), 4.54 (t, J = 6.6Hz, 2H), 4.04-4.01 (m, 2H), 4.00-3.93 (m, 4H), 3.91-3.80 (m, 6H), 3.73(dd, J = 11.3, 5.4 Hz, 2H), 3.63 (t, J = 9.4 Hz, 2H), 3.08 (t, J = 6.6Hz, 2H), 1.81-1.73 (m, 4H). 241 1-[2-[2,7-bis[2- (400 MHz, CD₃OD) δ 7.92(d, 733.21 [(2R, 3S, 4R, 5S, 6R)-3,4,5- J = 7.8 Hz, 2H), 7.85 (s, 2H),trihydroxy-6- 7.48 (d, J = 7.9 Hz, 2H), 4.95(hydroxymethyl)tetrahydropyran- (d, J = 4.2 Hz, 2H), 4.83-4.662-yl]ethynyl]spiro[fluorene-9,4′- (m, 6H), 4.47 (t, J = 6.0piperidine]-1′-yl]-2-oxo- Hz, 2H), 4.18 (s, 2H), 3.91-3.34ethyl]pyrrolidin-2-one (m, 18H), 2.25 (t, J = 8.0 Hz, 2H), 1.97 (d, J =7.6 Hz, 2H), 1.85 (s, 2H), 1.75 (s, 2H). 242 [2,7-bis[2-[(2R, 3S, 4R,5S, 6R)- (400 MHz, CD₃OD) δ 706.2 3,4,5-trihydroxy-6- 7.85-7.76 (m, 4H),7.52-7.48 (hydroxymethyl)tetrahydropyran- (m, 2H), 4.16-3.80 (m, 15H),2-yl]ethynyl]spiro[fluorene-9,4′- 3.73 (dd, J = 11.3, 5.5 Hz,piperidine]-1′-yl]-[(2S)- 2H), 3.63 (t, J = 9.4 Hz, 2H),tetrahydrofuran-2-yl]methanone 2.33-1.75 (m, 8H). C-1 sugar protons areunder the solvent peak 243 (5R)-5-[2,7-bis[2- (400 MHz, DMSO-d6) δ 719.2[(2R, 3S, 4R, 5S, 6R)-3,4,5- 7.97-7.89 (m, 3H), 7.78 (s, 1H),trihydroxy-6- 7.74 (s, 1H), 7.49 (d, J = 8.4(hydroxymethyl)tetrahydropyran- Hz, 2H), 4.98-4.92 (m, 2H),2-yl]ethynyl]spiro[fluorene-9,4′- 4.82-4.76 (m, 2H), 4.73 (d,piperidine]-1′-carbonyl]pyrrolidin- J = 2.1 Hz, 2H), 4.70 (dd, J = 2-one5.9 2.1 Hz, 2H), 4.65-4.59 (m, 1H), 4.51-4.44 (m, 2H), 3.97-3.34 (m,16H), 2.38-1.67 (m, 8H). 244 (5S)-5-[2,7-bis[2- (400 MHz, DMSO-d6) 719.2[(2R, 3S, 4R, 5S, 6R)-3,4,5- 8.16-8.13 (m, 1H), 8.00-7.88 trihydroxy-6-(m, 3H), 7.78 (s, 1H), 7.74 (s, (hydroxymethyl)tetrahydropyran- 1H),7.49 (d, J = 8.0 Hz, 2H), 2-yl]ethynyl]spiro[fluorene-9,4′- 5.02-4.89(m, 2H), 4.83-4.56 piperidine]-1′-carbonyl]pyrrolidin- (m, 7H),4.54-4.41 (m, 2-one 2H), 3.98-3.35 (m, 16H), 2.16-1.61 (m, 8H). 245[2,7-bis[2-[(2R, 3S, 4R, 5S, 6R)- (400 MHz, CD₃OD) δ 8.05 (s, 716.213,4,5-trihydroxy-6- 1H), 7.88-7.79 (m, 5H), 7.51(hydroxymethyl)tetrahydropyran- (dd, J = 7.9, 1.3 Hz, 2H),2-yl]ethynyl]spiro[fluorene-9,4′- 4.21-4.11 (m, 4H), 4.01 (dd,piperidine]-1′-yl]-(1 - J = 3.2, 2.1 Hz, 2H), 3.97-3.94methylpyrazol-4-yl)methanone (m, 2H), 3.93 (s, 3H), 3.91-3.80 (m, 4H),3.77-3.69 (m, 2H), 3.63 (t, J = 9.4 Hz, 2H), 1.98-1.90 (m, 4H).Differential Scanning Calorimetry of Compound 162Differential scanning calorimetry of Compound 162 crystalline form A canbe measured using the TA Instrument DSC Q200 (Asset V012390). A sample(1.02 mg) is weighed in a pre-punched pinhole aluminum hermetic pan andheated from ambient temperature to 350° C. at 10° C./min. The DSC resultseen in Figure A shows there is one endothermic peak observed, one at258° C. (onset temperature of 254° C., enthalpy 50.7 J/g).XRPD of Compound 162The XRPD can be recorded at room temperature in reflection mode usingBruker D8 Discover system (Asset Tag V012842) equipped with a sealedtube source and a Hi-Star area detector (Bruker AXS, Madison, Wis.). TheX-Ray generator operates at a tension of 40 kV and a current of 35 mA.The powder sample is placed on a Si zero-background wafer. Two framesare registered with an exposure time of 120 s each. The data aresubsequently integrated over the range of 3°-41° 2-theta with a stepsize of 0.02° and merged into one continuous pattern. Figure C shows theX-ray powder diffractogram of the sample. Representative XRPD peaks fromCompound 162:

Peak (2-Theta) Intensity 16.86 66.6 17.51 99.6 18.07 66.9 20.44 67.420.82 62.8 21.97 69.2 22.37 89.1 24.41 63 25.07 67.6 26.0 66.7 26.8762.8 31.89 59.1Thermo Analysis of Compound 202A thermal gravimetric analysis of Compound 202 was performed todetermine the percent weight loss as a function of time using the TAInstrument TGA Q500 (Asset V014840). A sample (1.29 mg) is added to apre-tared aluminum pan and heated from ambient temperature to 350° C. at10° C./min Weight loss ca. 2.2% was observed upon heating to 130° C.with degradation being observed at >250° C. The TGA result is shown inFigure D.XRPD of Compound 202The XRPD of Compound 202 was recorded at room temperature in reflectionmode using Bruker D8 Discover system (Asset Tag V012842) equipped with asealed tube source and a Hi-Star area detector (Bruker AXS, Madison,Wis.). The X-Ray generator was operating at a tension of 40 kV and acurrent of 35 mA. The powder sample was placed on a Si zero-backgroundwafer. Two frames were registered with an exposure time of 120 s each.The data were subsequently integrated over the range of 3.5°-39° 2-thetawith a step size of 0.02° and merged into one continuous pattern. FigureD shows the X-ray powder diffractogram of the sample.Representative XRPD Peaks from Compound 202

Peak (2-Theta) Intensity 7.42 38 9.39 65.8 14.29 91.6 14.9 100 16.2478.7Differential Scanning Calorimetry of Compound 202

Differential scanning calorimetry of Compound 202 crystalline form A canbe measured using the TA Instrument DSC Q2000 (Asset V012390). A sample(1.94 mg) is weighed in a pre-punched pinhole aluminum hermetic pan andrun with a modulation amplitude of +/−1° C./min using a ramp rate of 3°C./min to 300° C. The DSC result seen in figure E shows there is a glasstransition observed at ca. 143° C. followed by a melting endothermicpeak at 242° C. (onset temperature of 239.7° C., enthalpy 17.3 J/g).

Competitive Binding Assay

The first 177 amino acids of the FimH protein are expressed as a fusionprotein with thrombin in a pET21b plasmid in bacteria. This FimH proteinsequence contains the carbohydrate recognition domain (CRD) and shall betermed FimH-CRD. Following bacterial expression of the protein, theFimH-CRD protein is purified to homogeneity and the thrombin tag removedby protease cleavage. A competitive binding assay by fluorescencepolarization is performed using 5 nM of the Alexa 647 mannoside probeand 60 nM of the FimH-CRD. The samples are assayed in a low volume 384well microtiter plate in a final volume of 20 μl. The final assay bufferconditions are the following, 50 mM Tris-C1, ph 7.0, 100 mM NaCl, 1 mMEDTA, 5 mM β-mercaptoethanol, 0.05% BSA and 2.5% DMSO. Two assays areperformed for FimH, termed assay 1 or assay 2. The assay conditions arethe same for both assays except the following: assay 1 has compoundsprepared by manual dilution in a serial dilution factor with 12-pointdose response while assay 2 has compounds prepared by a robotics systemalso through a serial dilution factor (12 point dose response) andinitially prepared in duplicate in 384 well-Corning polypropylene roundbottom plates. The assay 2 plates have compound which is then frozen andmust be thawed prior to use. Initially the Alexa 647 probe and theFimH-CRD are added to the assay buffer and then 0.5 μl of test compound(assay 1 or 2) between 0.4 nM to 75 μM final concentration are added (12point titration with 3-fold serial dilution). Control wells for theAlexa 647 probe are prepared with the same conditions except for theaddition of the FimH-CRD protein. Plates are then incubated for 5 hrs atroom temperature in the dark and under humid conditions to preventdrying. Plates are read using the SpectraMax Paradigm multi-mode platereader and the appropriate fluorescent polarization detection cartridge(Alexa-647).

Alexa 647 mannoside probe is prepared using the similar procedurereported for FAM mannoside (Han, Z. et. al., 2010, J. Med. Chem., 53,4779) and is described in the scheme below.

To a blue colored stirred solution of(2S,3S,4S,5S,6R)-2-(4-aminobutoxy)-6-(hydroxymethyl)tetrahydropyran-3,4,5-triol(2.21 mg, 0.009 mmol) and the(2E)-2-[(2E,4E)-5-[3,3-dimethyl-5-sulfonato-1-(3-sulfonatopropyl)indol-1-ium-2-yl]penta-2,4-dienylidene]-3-[6-(2,5-dioxopyrrolidin-1-yl)oxy-6-oxo-hexyl]-3-methyl-1-(3-sulfonatopropyl)indoline-5-sulfonate(Potassium Ion (3)) (4.9 mg, 0.0044 mmol) in DMF (44 μL) is added Et₃N(5.4 mg, 7.0 μL, 0.053 mmol) at RT. The solution is stirred at roomtemperature over night, concentrated, dissolved in water and purified on12 g C-18 silica gel cartridge on Isolera system using acetonitrile inwater (0 to 40%, 10 CV) and followed by lyophilization to afford Alexa647 mannoside probe (3.3 mg, 34%) as deep blue solid.

The K_(d) values of the compounds are determined from dose responsecurves using twelve concentrations per compound in duplicate. Curves arefitted to data points using Fluorescence Polarization competitivedisplacement analysis, and Kds are interpolated from the resultingcurves using GraphPad Prism software, version 50.4 (GraphPad softwareInc., San Diego, Calif., USA). Table 1 below shows K_(d) values obtainedthrough both assay 1 and assay 2.

TABLE 1 Compound Assay 1 Assay 2 No. K_(d) (uM) K_(d) (uM)  1 <0.005,0.008, <0.005 0.013, 0.008  2 0.125, 0.062 NT  3 0.051, 0.04 NT  40.068, 0.096 NT  5 <0.005, <0.005 0.049, <0.005, <0.005  6 <0.005,<0.005 <0.005, 0.023  7 0.017, 0.011 NT  8 0.016, 0.019 NT  9 0.007,0.012 0.024 10 0.009, 0.015 NT 11 <0.005, 0.011 0.016 12 0.014, 0.018 NT13 0.017, 0.012 NT 14 0.011, 0.013 0.043, 0.006, 0.014 15 0.011, 0.015NT 16 <0.005, 0.006 0.026, <0.005, 0.009 17 <0.005, 0.006 NT 18 1.062,1.025 NT 19 0.006, <0.005 NT 20 0.058, 0.096 NT 21 <0.005, <0.005, 0.009NT 22 <0.005, 0.012 0.014, <0.005, 0.011 23 0.009, 0.012 0.025, <0.005,0.014 24 <0.005, <0.005 NT 25 <0.005, <0.005 0.021, 0.01, 0.012 26<0.005, <0.005 NT 27 <0.005, <0.005 NT 28 <0.005, <0.005 NT 29 0.006,0.008 NT 30 0.007, <0.005 NT 31 <0.005, <0.005 NT 32 <0.005, <0.005 NT33 <0.005, <0.005 NT 34 <0.005, 0.006 NT 35 <0.005, 12.357, 0.007 NT 360.006, 0.014, <0.005 NT 37 0.007, 0.007 0.018, <0.005, 0.012 38 <0.005,<0.005 <0.005, <0.005, 0.021 39 0.029, 0.046 NT 40 <0.005, 0.018 0.008,0.01 41 <0.005, <0.005 <0.005, <0.005 42 0.114, 0.377 0.14, 0.145 430.02, 0.027 NT 44 <0.005, <0.005 0.006 45 NT 0.707, 0.576, 0.558, 0.59146 NT 0.011, 0.009, 0.016, 0.014 47 NT 0.044, 0.037, 0.051, 0.036 48 NT0.079, 0.05, 0.063 49 NT 0.149, 0.087 50 NT 0.009, <0.005, <0.005 51 NT0.015, <0.005 52 NT <0.005, <0.005 53 NT <0.005, <0.005 54 NT 0.016,0.03 55 NT <0.005, 0.005 56 NT <0.005, <0.005 57 NT <0.005, <0.005 58 NT<0.005, <0.005 59 NT <0.005, <0.005 60 NT <0.005, <0.005 *NT means thecompound was not testedBacterial Binding AssayThe purpose of the Bacterial Binding Assay (BBA) is to determine theinhibition activity of selective FimH antagonists on the bacterialstrain LF82 binding to the glycoprotein BSA-(Mannose)₃.Below is a list of the Materials used to run the BBA are describedbelow.

-   -   1. LB broth: Supplier: Gibco, #10855    -   2. D-PBS: Supplier: Wisent, #311-425-CL    -   3. LB agar plates    -   4. 96-well black plate (high binding): Supplier: Costar, #3925    -   5. TopSeal™-A adhesive sealing films; Supplier PerkinElmer,        #6005185    -   6. Carbonate-bicarbonate buffer pH 9.6 tablets, Supplier:        Medicago, #09-8922-24    -   7. Water, Supplier: Gibco, #15230-162    -   8. Bovine serum albumin (BSA): Supplier: Sigma, # A-7888    -   9. (Man)3-BSA (α1-3, α1-6 Mannotriose-BSA, 1 mg), V-Labs, #        NGP1336, lot # HGDX37-169-1    -   10. Tween 20: Supplier: Sigma, # P9416    -   11. Bright-Glo Luciferase Assay System: Supplier: Promega, #        E2610    -   12. LF82/Luciferase strain: Invasive ability of an Escherichia        coli strain isolated from the ileal mucosa of a patient with        Crohn's disease. Boudeau J, Glasser A L, Masseret E, Joly B,        Darfeuille-Michaud A, Infect Immun. 1999, 67(9), 4499-509        Solutions and buffers used to run the BBA are described below.    -   1. 0.04M carbonate-bicarbonate buffer (coating buffer)    -   2. 40 μg/mL BSA-(Man)₃: Dissolve 1 mg of (Man)3-BSA in 25 mL of        water.    -   3. 4000 μg/mL BSA    -   4. 40 μg/mL BSA    -   5. 1 μg/mL BSA-(Man)₃: 150 μL of 40 μg/mL BSA-(Man)₃+5.85 mL of        40 μg/mL BSA    -   6. 0.5 μg/mL BSA-(Man)₃ in 0.02M carbonate-bicarbonate buffer.    -   7. 20 μg/mL BSA in 0.02M carbonate-bicarbonate buffer    -   8. Blocking buffer (2% BSA/DPBS): 1 g of BSA in 50 mL D-PBS    -   9. 2× binding buffer (0.2% BSA/D-PBS): 5 mL of blocking        buffer+45 mL D-PBS.    -   10. Washing buffer (D-PBS/0.01% Tween 20): 10 μL of Tween 20 in        100 mL D-PBS.    -   11. 1× Bright-Glo Luciferase substrate: Dilute 1:1 the        Bright-Glo Luciferase Assay System with D-PBS        The experimental protocol to run the BBA is described below.

Overnight culture of LF82/Luciferase strain: Into two Falcon 50 mLtubes, add 20 mL of LB+20 μL of 50 mg/mL Kanamycin and inoculate with aloop from glycerol stock of the LF82/Luciferase strain. Incubateovernight at 37° C. with no shaking.

Glycoprotein coating of 96-well plates: Add 100 μL/well of 0.5-2 μg/mLBSA-(Man)₃. 20 μg/mL BSA is used as the control background. Seal plateusing an adhesive sealing film and incubate overnight at roomtemperature. Wash the 96-well plate three times with 150 μL/well ofD-PBS, add 170 μL/well of blocking solution and incubate 45 min(minimum) at room temperature.

Preparation of bacterial suspension: Mix the two cultures tubes (40 mL)and perform a 1:10 dilution in LB (900 μl LB+100 μl culture. Measureoptical density (OD) of the bacterial cultures. OD1 ˜5×10⁸ cells/mL.Centrifuge LF82 culture for 20 min at 3500 rpm at room temperature.Re-suspend bacterial pellet in D-PBS and centrifuge again for 20 min at3500 rpm. Re-suspend bacterial pellet in D-PBS to obtain a bacterialconcentration of 2×10⁹ bacteria/mL. Dilute 1/10 in D-PBS to obtain afinal bacterial concentration of 2×10⁸ bacteria/mL (=107 bacteria/50μL). Perform 1/10 serial dilutions in LB of each bacterial suspension,plate 10 μL of dilutions on LB agar plates (final dilutions of 10⁻⁷) andincubate overnight at 37° C. and count CFUs to determine the actualbacteria density in the assay.

Bacterial binding assay: Add 147 μL 2× binding buffer to compound plate(containing 3 μL of compound). After blocking step is performed (atleast 45 min), wash plates three times with 200 μL/well of D-PBS. With a100 μl multichannel manual pipettor, add 504/well of compound diluted in2× binding buffer. With a 100 μL multichannel manual pipettor, add 50μL/well of bacterial suspension. Agitate at slow speed for 1 min andincubate 40-75 min at room temperature. Wash 5 times with 150 μL/well ofwashing buffer and then once with D-PBS. Add 100 μL/well of 1×Bright-Glo Luciferase substrate. Read luminescence by using the AnalystHT plate reader or the Trilux 1450 microbeta plate reader. Table 2 belowprovides IC50 data for compounds 1-245 in the bacterial binding assay.

TABLE 2 Bacterial Binding Assay Bacterial Binding Assay Compound IC₅₀(μM)   1 5.267   2 4.95   3 NT   4 NT   5 0.58   6 0.095   7 NT   8 NT  9 NT  10 NT  11 NT  12 NT  13 NT  14 4.10  15 NT  16 1.51  17 NT  18NT  19 0.43  20 NT  21 NT  22 1.60  23 5.80  24 0.65  25 1.40  26 0.85 27 0.80  28 0.77  29 1.30  30 0.30  31 NT  32 0.34  33 0.39  34 NT  35NT  36 NT  37 1.65  38 0.41  39 NT  40 0.043  41 0.55  42 NT  43 NT  440.48  45 NT  46 NT  47 NT  48 NT  49 NT  50 0.95  51 0.83  52 0.66  530.055  54 1.15  55 0.21  56 0.020  57 0.027  58 0.017  59 0.0082  600.013  62 0.067  63 0.112  64 0.015  65 0.055  66 0.034  67 0.040  680.0077  69 0.066  70 0.015  71 0.028  72 0.018  73 0.021  74 0.0081  750.076  76 0.046  77 0.0040  78 0.063  79 0.037  80 0.011  81 0.009  820.018  83 0.0038  84 0.315  85 0.16  86 0.022  87 0.017  88 0.132  890.082  90 0.036  91 0.058  92 0.036  93 0.088  94 0.020  95 0.038  960.066  97 0.024  98 0.012  99 0.145 100 0.028 101 0.021 102 0.024 1030.0074 104 0.034 105 0.046 106 0.0079 107 0.013 108 0.028 109 0.020 1100.00089 111 0.0021 112 0.017 113 0.0055 114 0.0012 115 0.0039 116 0.0081117 0.023 118 0.022 119 0.014 120 0.0012 121 0.012 122 0.029 123 0.041124 0.027 125 0.054 126 0.124 127 0.043 128 0.022 129 1.245 130 0.067131 0.128 132 0.052 133 0.028 134 0.044 135 0.58 136 0.145 137 0.70 1380.031 139 0.046 140 0.353 141 0.34 142 0.091 143 0.24 144 0.235 145 0.43146 0.165 147 0.134 148 0.13 149 0.027 150 0.071 151 0.185 152 0.031 1530.092 154 0.044 155 0.022 156 0.018 157 0.010 158 0.011 159 0.046 1600.017 161 0.012 162 0.00011 163 0.018 164 0.035 165 0.0024 166 0.003 1670.016 168 0.0031 169 0.00066 170 0.0075 171 0.013 172 0.188 173 0.0093174 0.0047 175 0.0065 176 0.010 177 0.00066 178 0.0031 179 0.0071 1800.0055 181 0.010 182 0.0014 183 0.00076 184 0.00083 185 0.0011 1860.00028 187 0.0010 188 0.0043 189 0.0078 190 0.0011 191 0.00036 1920.0032 193 0.00027 194 0.00027 195 0.018 196 0.028 197 0.00083 198 0.024199 0.00021 200 0.00005 201 0.026 202 0.00007 203 0.00004 204 0.00080205 0.00039 206 0.00008 207 0.00005 208 0.00003 209 0.00007 210 0.00016211 0.0022 212 0.0049 213 0.0037 214 0.00045 215 0.0050 216 0.0099 2170.020 218 0.00009 219 0.0038 220 0.00003 221 0.00004 222 0.00008 2230.00006 224 0.00008 225 0.0012 226 0.00006 227 0.00006 228 0.00044 2290.00035 230 0.00016 231 0.00008 232 0.00004 233 0.00005 234 0.00004 2350.00003 236 0.00050 237 0.00022 238 0.00009 239 0.00035 240 0.00005 2410.00003 242 0.00018 243 0.00022Mouse Model of Inflammatory Bowel Disease (IBD):

Transgenic humanized-CEACAM6 mouse model may be used to test thecompounds of the invention (Carvalho F A et al. (2009) J Exp Med.September 28; 206(10):2179-89). The Transgenic humanized-CEACAM6 miceare infected as described in Carvalho et al. The infected mice can thentreated with compounds of the present invention.

While we have described a number of embodiments of this invention, it isapparent that our basic examples may be altered to provide otherembodiments that utilize the compounds, methods, and processes of thisinvention. Therefore, it will be appreciated that the scope of thisinvention is to be defined by the appended claims rather than by thespecific embodiments that have been represented by way of exampleherein.

What is claimed is:
 1. A compound of Formula I, or a pharmaceuticallyacceptable salt thereof:

wherein Z is

Ring H is an optionally substituted 5-6 membered aromatic monocyclicring optionally having 1-4 heteroatoms selected from nitrogen, orsulfur; an 8-12 membered aromatic bicyclic ring optionally having 1-6heteroatoms selected from oxygen, nitrogen, or sulfur; or a 10-14membered aromatic tricyclic ring optionally having 1-6 heteroatomsselected from oxygen, nitrogen, or sulfur; L² is —X²

Y²—wherein X² is a C₁aliphatic or —C(O)— and Y² is C₁-C₁₀aliphaticwherein up to two methylene units of the C₁-C₁₀aliphatic are optionallyreplaced with —C(O)—, NH, or N(C₁-C₆aliphatic); L² is optionallysubstituted with 1-3 halo; L³ is C₁-C₁₂aliphatic wherein up to threemethylene units of the C₁-C₁₂aliphatic are optionally replaced with—C(O)—, NH, or N(C₁-C₆aliphatic); L³ is optionally substituted with 1-3halo; each J^(H) is independently halogen, —CN, —NO₂, X^(J), Q^(J), orX^(J)-Q^(J); or two J^(H) groups bound to the same carbon atom, togetherwith the carbon atom to which they are bound, optionally form —C═N—OH,—C(O)—, or Ring HH; Ring HH is a 3-8 membered saturated monocyclic ringhaving 0-2 heteroatoms selected from oxygen, nitrogen, or sulfur;optionally substituted with 1-4 occurrences of J^(HH); J^(HH) is halo,CN, oxo, X^(J), Q^(J), or X^(J)-Q^(J); X^(J) is a C₁-C₁₀ aliphatic,wherein up to 4 methylene units of the C₁-C₁₀ aliphatic are optionallyreplaced with —O—, —NH, N(C₁C₆aliphatic), —S—, —C(O)—, —C(═NOH)—,—S(O)—,—S(O)₂—, P, or P(O); X^(J) is optionally substituted with 0-6occurrences of halo, OH, or C₁₋₄alkyl; or optionally substituted with0-1 occurrences of CN and; Q^(J) is a 3-7 membered monocyclic saturated,fully unsaturated, partially unsaturated, or aromatic ring optionallyhaving 1-4 heteroatoms selected from oxygen, nitrogen, or sulfur; or an8-12 membered saturated, fully unsaturated, partially unsaturated, oraromatic ring optionally having 1-6 heteroatoms selected from oxygen,nitrogen, or sulfur; wherein each Q^(J) is optionally substituted with1-6 occurrences of halo, oxo, CN, or C₁₋₆alkyl, wherein up to 2methylene units of said C₁₋₆alkyl are optionally replaced with —O—, —NH,N(C₁-C₆aliphatic), —S—, —C(O)—, —S(O)—, or —S(O)₂—.
 2. The compound ofclaim 1, wherein Ring H, is phenyl, napthyl, or a 5-6 memberedheteroaryl having 1-4 heteroatoms selected from oxygen, nitrogen, orsulfur.
 3. The compound of claim 1, wherein Z is

and J^(H) is halo, CN, NO₂, phenyl, or C₁₋₁₀aliphatic wherein up to 3methylene units are optionally replaced with O, NH, N(C₁₋₄alkyl), S,C(O), SO, or SO₂; wherein said J^(H) is optionally substituted with 1-3occurrences of CN, halo or phenyl.
 4. The compound of claim 3 wherein L²is C₁₋₆aliphatic or (C₁₋₄aliphatic)-C(O)NH—; L³ is C₁₋₆aliphatic or—NHC(O)—(C₁₋₄aliphatic)-; Ring H is phenyl or naphthyl; and J^(H) ishalo, CN, NO₂, C₁₋₆aliphatic, —OC₁₋₆aliphatic, or C(O)O(C₁₋₆aliphatic);wherein said J^(H) is optionally substituted with 1-3 occurrences ofhalo.
 5. The compound of claim 1, wherein L² and L³ are eachindependently C₁-C₄ alkenyl or C₁-C₄ alkynyl.
 6. The compound of claim1, having formula IB:


7. The compound of claim 6, wherein L² and L³ are bonded to the mannosering via a carbon atom.
 8. The compound of claim 7, wherein L² and L³are each independently C₁-C₆ alkenyl or C₁-C₆ alkynyl.
 9. The compoundof claim 8, wherein at least one of L² and L³ is —C≡C—.
 10. The compoundof claim 1, having formula ID:


11. The compound of claim 10, wherein Ring H is an optionallysubstituted 5-6 membered monocyclic aromatic ring optionally having 1-4heteroatoms selected from nitrogen, or sulfur; or an 8-12 memberedbicyclic aromatic ring optionally having 1-6 heteroatoms selected fromoxygen, nitrogen, or sulfur; or a 10-14 tricyclic aromatic ringoptionally having 1-6 heteroatoms selected from oxygen, nitrogen, orsulfur.
 12. The compound of claim 11, wherein Ring H is optionallysubstituted phenyl, naphthyl, thienyl, isoxazolyl, pyridinyl, pyrazinyl,indolyl, indazolyl, thienylthiophenyl, quinolinyl, quinazolinyl,benzothiadiazolyl, or fluorenyl.
 13. The compound of claim 10, whereinRing H, together with J^(H) and J^(HH), is selected from the following:


14. The compound of claim 10, wherein J^(H) is halogen, oxo, CN, Q^(J),or X^(J)-Q^(J); wherein X^(J) is C₁-C₁₀ aliphatic, wherein up to 4methylene units of the C₁-C₁₀ aliphatic are optionally replaced with—O—, —NH, N(C₁-C₆aliphatic), —S—, —C(O)—, —S(O)—, —S(O)₂—; Q^(J) isphenyl; and J^(H) is optionally substituted with 0-3 occurrences of haloor 0-1 occurrences of CN.
 15. The compound of claim 14, wherein J^(H) ishalogen, CN, —C(CH₃)₂CN, C₃₋₆cycloalkyl, phenyl, —O—CH₂phenyl, orC₁₋₆alkyl wherein up to one methylene unit is optionally replaced with—O—, —S—, —NH—, —N(C₁₋₆alkyl)-, or —C(O)—; wherein said J^(H) issubstituted with 0-3 halo or 0-1 CN.
 16. The compound of claim 10,wherein Ring H is optionally substituted phenyl or naphthyl.
 17. Thecompound of claim 16, wherein Ring H is phenyl and J^(H) is halo, CN,—C(CH₃)₂CN, C₃₋₆cycloalkyl, phenyl, CH₂phenyl, —O—CH₂phenyl, orC₁₋₆alkyl wherein up to one methylene unit is optionally replaced with—O—, —S—, —NH—, —N(C₁₋₆alkyl)-, or —C(O)—; wherein said J^(H) issubstituted with 0-3 halo or 0-1 CN.
 18. The compound of claim 1,represented by a structural formula selected from the group consistingof:

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or a pharmaceutically acceptable salt thereof.
 19. The compound of claim18, wherein the compound is compound 53:


20. A composition comprising the compound of claim 1, or apharmaceutically acceptable salt thereof, and a pharmaceuticallyacceptable carrier, adjuvant, or vehicle.